Pyruvate kinase M2 modulators, therapeutic compositions and related methods of use

ABSTRACT

Compositions comprising compounds that modulate pyruvate kinase M2 (PKM2) are described herein. Also described herein are methods of using the compounds that modulate PKM2 in the treatment of cancer.

CLAIM OF PRIORITY

This application is a continuation of U.S. Ser. No. 14/277,957, filedMay 15, 2014, which is a divisional of U.S. Ser. No. 13/267,494, filedOct. 6, 2011, now U.S. Pat. No. 8,742,119, issued Jun. 3, 2014, which isa continuation of International Application No. PCT/US2010/030139, filedApr. 6, 2010, which claims priority from U.S. Ser. No. 61/233,470, filedAug. 12, 2009, and U.S. Ser. No. 61/167,017, filed Apr. 6, 2009, each ofwhich is incorporated herein by reference in its entirety.

BACKGROUND OF INVENTION

Cancer cells rely primarily on glycolysis to generate cellular energyand biochemical intermediates for biosynthesis of lipids andnucleotides, while the majority of “normal” cells in adult tissuesutilize aerobic respiration. This fundamental difference in cellularmetabolism between cancer cells and normal cells, termed the WarburgEffect, has been exploited for diagnostic purposes, but has not yet beenexploited for therapeutic benefit.

Pyruvate kinase (PK) is a metabolic enzyme that convertsphosphoenolpyruvate to pyruvate during glycolysis. Four PK isoformsexist in mammals: the L and R isoforms are expressed in liver and redblood cells, the M1 isoform is expressed in most adult tissues, and theM2 isoform is a splice variant of M1 expressed during embryonicdevelopment. All tumor cells exclusively express the embryonic M2isoform. A well-known difference between the M1 and M2 isoforms of PK isthat M2 is a low-activity enzyme that relies on allosteric activation bythe upstream glycolytic intermediate, fructose-1,6-bisphosphate (FBP),whereas M1 is a constitutively active enzyme.

All tumor cells exclusively express the embryonic M2 isoform of pyruvatekinase, suggesting PKM2 as a potential target for cancer therapy. PKM2is also expressed in adipose tissue and activated T-cells. Thus, themodulation (e.g., inhibition or activation) of PKM2 may be effective inthe treatment of, e.g., obesity, diabetes, autoimmune conditions, andproliferation-dependent diseases, e.g., benign prostatic hyperplasia(BPH). Current modulators (e.g., inhibitors) of pyruvate kinase are notselective, making it difficult to treat disease related to pyruvatekinase function.

Furthermore, phosphotyrosine peptide binding to PKM2 leads to adissociation of FBP from PKM2 and conformational changes of PKM2 from anactive, tetrameric form to an inactive form. Compounds that bind to PKM2and lock the enzyme in the active confirmation will lead to the loss ofallosteric control of PKM2 needed for shunting biochemical intermediatesfrom glycolysis into biosynthesis of nucleotides and lipids. Thus, theactivation of PKM2 can also inhibit the growth and proliferation ofcancer cells, activated immune cells, and fat cells.

There is a continuing need for novel treatments of diseases such ascancer, diabetes, obesity, autoimmune conditions,proliferation-dependent diseases (e.g., BPH), and other diseases relatedto the function of pyruvate kinase (e.g., PKM2).

SUMMARY OF INVENTION

Described herein are compounds that modulate pyruvate kinase M2 (PKM2)and pharmaceutically acceptable salts, solvates, and hydrates thereof,for example, compounds that modulate PKM2. This invention also providescompositions and pharmaceutical kits comprising a compound of thisinvention and the use of such compositions and kits in methods oftreating diseases and conditions that are related to pyruvate kinasefunction (e.g., PKM2 function), including, e.g., cancer, diabetes,obesity, autoimmune disorders, and benign prostatic hyperplasia (BPH).

In one aspect, the present invention features a compound orpharmaceutically acceptable salt thereof of formula (I):

wherein

-   -   X¹ is N or CE;    -   X² is N or CD;    -   X³ is N or CB;    -   X⁴ is N or CA;    -   Y¹, Y², Y³ and Y⁴ are each independently selected from N and        CR¹;    -   A, B, D and E are each independently selected from H, R³ and        —SO₂—NR⁴R⁵;    -   wherein at least one of X¹, X², X³, X⁴, Y¹, Y², Y³ and Y⁴ is N;        and at least one of X¹, X², X³, X⁴, is C—SO₂—NR⁴R⁵;    -   each R⁴ is independently selected from C₁₋₈ alkyl, aryl and        heteroaryl, each of which is substituted with n occurrences of        R²;    -   each R⁵ is independently hydrogen or C₁₋₈ alkyl;    -   each R¹ is independently selected from hydrogen, C₁₋₈ alkyl,        C₁₋₈ terminal alkynyl, C₁₋₈ alkoxy, halogen, haloalkyl and        haloalkoxy;    -   each R² is independently selected from halo, haloalkyl, C₁₋₄        alkyl, C₂₋₄ alkenyl, C₂₋₄ alknynyl, aryl, heteroaryl, aralkyl,        heteroaralkyl, cyano, —OR^(a), —COOR^(b) and —CONR^(c)R^(c′);        wherein two R², together with the carbons to which they are        attached, may form an optionally substituted ring, each of which        can be further substituted;    -   each R³ is independently selected from C₁₋₈ alkyl, —OR^(a),        halogen, haloalkyl, haloalkoxy and optionally substituted        heteroaryl;    -   each R^(a) is independently selected from alkyl, haloalkyl,        optionally substituted heteroaryl and optionally substituted        heterocyclyl;    -   each R^(b) is independently alkyl; and    -   each R^(c) is independently selected from hydrogen and alkyl;        and    -   n is 0, 1, 2 or 3.

In some embodiments, one of X¹, X², X³, X⁴, is C—SO₂—NR⁴R⁵;

In some embodiments, X⁴ is A and A is —SO₂—NR⁴R⁵. In some embodiments,X³ is B and B is —SO₂—NR⁴R⁵. In some embodiments, X² is D and D is—SO₂—NR⁴R⁵. In some embodiments, X¹ is CE and E is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ia):

wherein A, B, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ib):

wherein A, B, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ic):

wherein A, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or D is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Id):

wherein A, D, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ie):

wherein A, B, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (If):

wherein A, B, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ig):

wherein A, B, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ih):

wherein A, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ij):

wherein A, D, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ik):

wherein A, B, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Im):

wherein A, B, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (In):

wherein A, B, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Io):

wherein A, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ip):

wherein A, D, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iq):

wherein A, B, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ir):

wherein A, B, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Is):

wherein A, B, D, E, R¹, R², R³, R⁴, R⁵, m and n are as defined informula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (It):

wherein A, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iu):

wherein A, D, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iv):

wherein A, B, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iw):

wherein A, B, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A or B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ix):

wherein A, B, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iy):

wherein A, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iz):

wherein A, D, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iaa):

wherein A, B, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Ibb):

wherein A, B, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Icc)

wherein A, B, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, B is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Idd):

wherein A, D, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iee):

wherein A, D, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, the compound is a compound of formula (Iff):

wherein A, B, E, R¹, R², R³, R⁴, R⁵ and n are as defined in formula (I).

In some embodiments, A is —SO₂—NR⁴R⁵.

In some embodiments, B is —SO₂—NR⁴R⁵.

In one aspect, the invention features a pharmaceutical compositioncomprising a compound of formula (I) as described herein, or apharmaceutically acceptable salt thereof.

In one aspect, the invention features a method of treating a disorderdescribed herein (e.g., cancer) comprising administering to a subject acompound of formula (I) as described herein or a pharmaceuticallyacceptable salt thereof.

In one aspect, the invention features a method of preventing (e.g.,preventing the onset of at least one symptom) or delaying the onset of adisorder as described herein (e.g., cancer) comprising administering toa subject a compound of formula (I) as described herein or apharmaceutically acceptable salt thereof.

In one aspect, the present invention features a compound orpharmaceutically acceptable salt thereof of formula (II):

wherein

A, B, D and E are each independently selected from H, —SO₂—NR⁴R⁵ and R³;wherein at least one of A, B, D, or E is —SO₂—NR⁴R⁵;

Y, Y², Y³ and Y⁴ are each independently selected from N and CR¹, whereinat least one of Y¹, Y², Y³ and Y⁴ are N;

each R⁴ is independently selected from C₁₋₈ alkyl, aryl and heteroaryl,each of which is substituted with n occurrences of R²;

each R⁵ is independently hydrogen or C₁₋₈ alkyl;

each R¹ is independently selected from hydrogen, C₁₋₈ alkyl, C₁₋₈terminal alkynyl, C₁₋₈ alkoxy, halogen, haloalkyl and haloalkoxy;

each R² is independently selected from halo, haloalkyl, C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alknynyl, aryl, heteroaryl, aralkyl, heteroaralkyl, cyano,—OR^(a), —COOR^(b) and —CONR^(c)R^(c′); wherein two R², together withthe carbons to which they are attached, may form an optionallysubstituted ring, each of which can be further substituted;

each R³ is independently selected from C₁₋₈ alkyl, —OR^(a), halogen,haloalkyl, haloalkoxy and optionally substituted heteroaryl;

each R^(a) is independently selected from alkyl, haloalkyl, optionallysubstituted heteroaryl and optionally substituted heterocyclyl;

each R^(b) is independently alkyl; and

each R^(c) is independently selected from hydrogen and alkyl; and

n is 0, 1, 2 or 3.

In some embodiments, at least one of Y¹, Y², Y³ and Y⁴ is N. In someembodiments, at least one of Y¹, Y², Y³ and Y⁴ are CH. In someembodiments, Y¹ is N. In some embodiments, Y³ is N.

In some embodiments, each R¹ is independently hydrogen.

In some embodiments, the invention features a compound of formula (IIa):

wherein n, B, D, E, R¹, R², R³, R⁴ and R⁵ are defined as above.

In some embodiments, the invention features a compound of formula (IIb):

wherein n, B, D, E, R¹, R², R³, R⁴ and R⁵ are defined as above.

In some embodiments, B, D and E are each independently selected from H.

In some embodiments, R⁵ is hydrogen.

In some embodiments, each R¹ is independently hydrogen. In someembodiments, each R¹ is independently selected from C₁₋₈ alkyl, halogenor haloalkyl. In some embodiments, each R¹ is independently selectedfrom halogen or haloalkyl. In some embodiments, each R¹ is independentlyselected from halogen (e.g., chlorine or fluorine). In some embodiments,each R¹ is independently haloalkyl (e.g., trifluoroalkyl).

In some embodiments, R⁴ is selected from aryl or heteroaryl. In someembodiments, R⁴ is aryl substituted with n occurrences of R². In someembodiments, R⁴ is C₅₋₈ monocyclic aryl or C₈₋₁₄ bicyclic aryl. In someembodiments, R⁴ is C₅₋₈ monocyclic aryl (e.g., optionally substitutedphenyl). In some embodiments, R⁴ is phenyl substituted with noccurrences of R².

In some embodiments, n is 0. In some embodiments, n is 1.

In some embodiments, R² is halo, C₁₋₄ alkyl or haloalkyl, each of whichcan be further substituted.

In some embodiments, R² is C₁₋₄ alkyl (e.g., ethyl). In someembodiments, R² is halo (e.g., chloro). In some embodiments, R² ishaloalkyl (e.g., trifluoromethyl).

In some embodiments, n is 2. In some embodiments, both R² are C₁₋₄ alkyl(e.g., methyl). In some embodiments, n is 2. In some embodiments, bothR² are halo (e.g., fluoro or chloro). In some embodiments, n is 2. Insome embodiments, one R² is haloalkyl (e.g., trifluoroalkyl) and theother R² is —OR^(a). In some embodiments, R^(a) is alkyl (e.g., methylor ethyl). In some embodiments, n is 2. In some embodiments, one R² ishalo (e.g., fluoro or chloro) and the other R² is C₁₋₄ alkyl (e.g.,methyl or ethyl).

In some embodiments, n is 2. In some embodiments, two R², together withthe carbon atoms to which they are attached, form a 5-memberedheterocyclic ring. In some embodiments, two R², together with the phenylring to which they are attached, form the following structure:

In some embodiments, n is 3. In some embodiments, all R² are halo (e.g.,fluoro or chloro).

In another aspect, the invention features a pharmaceutical compositioncomprising a compound selected from Formula (II), (IIa) or (IIb) asdescribed herein or a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of treating adisorder described herein (e.g., cancer) comprising administering to asubject a compound of formula (II), (IIa) or (IIb) as described hereinor a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of preventing (e.g.,preventing the onset of at least one symptom) or delaying the onset of adisorder as described herein (e.g., cancer) comprising administering toa subject a compound of formula (II), (IIa) or (IIb) as described hereinor a pharmaceutically acceptable salt thereof.

In one aspect, the present invention features a compound orpharmaceutically acceptable salt thereof of formula (III):

wherein

-   -   X¹ is N or CE;    -   X² is N or CD;    -   X³ is N or CB;    -   X⁴ is N or CA, wherein at least one of X¹, X², X³ and X⁴ is N        and at least one of X¹, X², X³, X⁴, is C—SO₂—NR⁴R⁵;    -   A, B, D and E are each independently selected from H, R³ and        —SO₂—NR⁴R⁵;    -   Y² is selected from N and CR¹;    -   each R⁴ is independently selected from C₁₋₈ alkyl, aryl and        heteroaryl, each of which is substituted with n occurrences of        R²;    -   R⁵ is hydrogen or C₁₋₈ alkyl;    -   each R¹ is independently selected from hydrogen, C₁₋₈ alkyl,        C₁₋₈ terminal alkynyl, C₁₋₈ alkoxy, halogen, haloalkyl and        haloalkoxy;    -   each R² is independently selected from halo, haloalkyl, C₁₋₄        alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, aryl, heteroaryl, aralkyl,        heteroaralkyl, cyano, —OR^(a), —COOR^(b) and —CONR^(c)R^(c′);        wherein two R², together with the carbons to which they are        attached, may form an optionally substituted ring, each of which        can be further substituted;    -   each R³ is independently selected from C₁₋₈ alkyl, —OR^(a),        halogen, haloalkyl, haloalkoxy and optionally substituted        heteroaryl;    -   each R^(a) is independently selected from alkyl, haloalkyl,        optionally substituted heteroaryl and optionally substituted        heterocyclyl;    -   each R^(b) is independently alkyl; and    -   each R^(c) is independently selected from hydrogen and alkyl;        and    -   n is 0, 1, 2 or 3.

In some embodiments, the invention features a compound of formula(IIIa):

wherein n, B, D, E, R¹, R⁴, R² and R⁵ are defined as in formula (III).

In some embodiments, the invention features a compound of formula(IIIb):

wherein n, B, E, R¹, R⁴, R² and R⁵ are defined as in formula (III).

In some embodiments, the invention features a compound of formula(IIIc):

wherein n, B, D, R¹, R⁴, R² and R⁵ are defined as in formula (III).

In some embodiments, B and E are each independently selected from H.

In some embodiments, R⁵ is hydrogen.

In some embodiments, each R¹ is independently hydrogen. In someembodiments, each R¹ is independently selected from C₁₋₈ alkyl, halogenor haloalkyl. In some embodiments, each R¹ is independently selectedfrom halogen or haloalkyl. In some embodiments, each R¹ is independentlyselected from halogen (e.g., chlorine or fluorine). In some embodiments,each R¹ is independently haloalkyl (e.g., trifluoroalkyl).

In some embodiments, R⁴ is selected from aryl or heteroaryl. In someembodiments, R⁴ is aryl substituted with n occurrences of R². In someembodiments, R⁴ is C₅₋₈ monocyclic aryl or C₈₋₁₄ bicyclic aryl. In someembodiments, R⁴ is C₅₋₈ monocyclic aryl (e.g., optionally substitutedphenyl). In some embodiments, R⁴ is phenyl substituted with noccurrences of R².

In some embodiments, n is 0. In some embodiments, n is 1.

In some embodiments, R² is halo, C₁₋₄ alkyl or haloalkyl, each of whichcan be further substituted.

In some embodiments, R² is C₁₋₄ alkyl (e.g., methyl or ethyl). In someembodiments, R² is halo (e.g., chloro).

In some embodiments, n is 2. In some embodiments, both R² are C₁₋₄ alkyl(e.g., methyl or ethyl). In some embodiments, n is 2. In someembodiments, both R² are halo (e.g., fluoro or chloro). In someembodiments, n is 2. In some embodiments, one R² is haloalkyl (e.g.,trifluoroalkyl) and the other R² is —OR^(a). In some embodiments, R^(a)is alkyl (e.g., methyl or ethyl). In some embodiments, n is 2. In someembodiments, one R² is C₁₋₄ alkyl (e.g., methyl or ethyl) and the otherR² is halo (e.g., fluoro or chloro).

In some embodiments, n is 2. In some embodiments, two R², together withthe carbon atoms to which they are attached, form a 5-memberedheterocyclic ring. In some embodiments, two R², together with the phenylring to which they are attached, form the following structure:

In some embodiments, n is 3. In some embodiments, all R² are halo (e.g.,fluoro or chloro).

In another aspect, the invention features a pharmaceutical compositioncomprising a compound selected from formula (III), (IIIa), (IIIb) or(IIIc) as described herein or a pharmaceutically acceptable saltthereof.

In another aspect, the invention features a method of treating adisorder described herein (e.g., cancer) comprising administering to asubject a compound of formula (III), (IIIa), (IIIb) or (IIIc) asdescribed herein or a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of preventing (e.g.,preventing the onset of at least one symptom) or delaying the onset of adisorder described here (e.g., cancer) comprising administering to asubject a compound of formula (III), (IIIa), (IIIb) or (IIIc) asdescribed herein or a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a compound or pharmaceuticallyacceptable salt thereof selected from the following formula:

wherein

-   -   n is 0, 1, 2 or 3;    -   X¹ is N or CE;    -   X² is N or CD;    -   X³ is N or CB;    -   X⁴ is N or CA, wherein at least one of X¹, X², X³ and X⁴ is N;        and at least one of X¹, X², X³, X⁴, is C—SO₂—NR⁴R⁵;    -   A, B, D and E are each independently selected from H and        —SO₂—NR⁴R⁵;    -   each R⁴ is independently selected from C₁₋₅ alkyl, aryl and        heteroaryl, each of which is substituted with n occurrences of        R²;    -   each R⁵ is independently hydrogen or C₁₋₈ alkyl;    -   each R¹ is independently selected from hydrogen, C₁₋₅ alkyl,        C₁₋₅ alkoxy, halogen, haloalkyl and haloalkoxy;    -   each R² is independently selected from halo, haloalkyl,        C₁₋₄alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynylheteroaryl, aryl, aralkyl,        heteroaralkyl, cyano, —OR^(a), —COOR^(b) and —CONR^(c)R^(c′);        wherein two R², together with the carbons to which they are        attached, may form an optionally substituted ring, each of which        can be further substituted;    -   each R³ is independently selected from C₁₋₅ alkyl, —OR^(a),        halogen, haloalkyl, haloalkoxy or optionally substituted        heteroaryl;    -   R^(a) is independently selected from alkyl, haloalkyl,        optionally substituted heteroaryl and optionally substituted        heterocyclyl;    -   each R^(b) is independently alkyl; and    -   each R^(c) is independently selected from hydrogen and alkyl.

In some embodiments, at least one of X³ and X⁴ are CH.

In some embodiments, at least one of A, B, D and E are H. In someembodiments, at least one of A, B, D and E are —SO₂—NH—R⁴. In someembodiments, A is —SO₂—NH—R⁴. In some embodiments, B is —SO₂—NH—R⁴.

In some embodiments, the invention features a compound of formula (IVa):

wherein n, B, E, R¹, R⁴, R³, R² and R⁵ are defined as above.

In some embodiments, the invention features a compound of formula (IVb):

wherein n, A, D, E, R¹, R⁴, R³, R², and R⁵ are defined as above.

In some embodiments, A, and D are H. In some embodiments, B and E are H.

In some embodiments, R⁵ is hydrogen.

In some embodiments, each R¹ is independently H. In some embodiments,each R¹ is independently selected from C₁₋₈ alkyl, halogen or haloalkyl.In some embodiments, each R¹ is independently selected from halogen orhaloalkyl. In some embodiments, each R¹ is independently selected fromhalogen (e.g., chlorine or fluorine). In some embodiments, each R¹ isindependently haloalkyl (e.g., trifluoroalkyl).

In some embodiments, R⁴ is selected from aryl or heteroaryl. In someembodiments, R⁴ is aryl substituted with n occurrences of R². In someembodiments, R⁴ is C₅₋₈ monocyclic aryl or C₈₋₁₄ bicyclic aryl. In someembodiments, R⁴ is C₅₋₈ monocyclic aryl (e.g., phenyl). In someembodiments, R⁴ is phenyl substituted with n occurrences of R². In someembodiments, R⁴ is C₈₋₁₄ bicyclic aryl (e.g., napthyl). In someembodiments, R⁴ is a 5-8 membered heteroaryl or a 8-14 memberedheteroaryl. In some embodiments, R⁴ is a 8-14 membered heteroaryl (e.g.,5-quinolyl or 6-quinolyl). In some embodiments, R⁴ is quinolyl (e.g.,5-quinolyl or 6-quinolyl) substituted with n occurrences of R².

In some embodiments, n is 0. In some embodiments, n is 1.

In some embodiments, R² is selected from halo, C₁₋₄ alkyl, cyano,haloalkyl, —OR^(a) or two R², taken together with the carbon atoms towhich they are attached form an optionally substituted ring, each ofwhich can be further substituted.

In some embodiments, R² is halo (e.g., chloro or fluoro). In someembodiments, R² is C₁₋₄ alkyl (e.g., methyl or ethyl). In someembodiments, R² is cyano. In some embodiments, R² is haloalkyl (e.g.,trifluoromethyl). In some embodiments, R² is —OR^(a). In someembodiments, R^(a) is alkyl (e.g., methyl).

In some embodiments, n is 2. In some embodiments, both R² are C₁₋₄ alkyl(e.g., methyl). In some embodiments, n is 2. In some embodiments, bothR² are halo (e.g., fluoro or chloro). In some embodiments, n is 2. Insome embodiments, one R² is C₁₋₄ alkyl and the other is halo (e.g.,methyl and chloro or methyl and fluoro). In some embodiments, n is 2. Insome embodiments, both R² are haloalkyl (e.g., trifluoromethyl). In someembodiments, n is 2. In some embodiments, both R² are —OR^(a). In someembodiments, both R^(a) are alkyl (e.g, methyl). In some embodiments, nis 2. In some embodiments, one R² is haloalkyl (e.g., trifluoromethyl)and one is —OR^(a). In some embodiments, R^(a) is alkyl (e.g., methyl).

In some embodiments, n is 2. In some embodiments, two R², taken togetherwith the carbon atoms to which they are attached, form a 6-memberedheterocyclic ring. In some embodiments, two R², taken together with thephenyl ring to which they are attached, for the following structure:

In some embodiments, n is 3. In some embodiments, three R² are halo(e.g., fluoro).

In another aspect, the invention features a pharmaceutical compositioncomprising a compound selected from formula (IV), (IVa) or (IVb) asdescribed herein or a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of treating adisorder described herein (e.g., cancer) comprising administering to asubject a compound of formula (IV), (IVa) or (IVb) as described hereinor a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of preventing (e.g.,preventing the onset of at least one symptom) or delaying the onset of adisorder described here (e.g., cancer) comprising administering to asubject a compound of formula (IV), (IVa) or (IVb) as described hereinor a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a pharmaceutical compositioncomprising a compound of formula (V):

wherein

B, D and E are each independently selected from H and R³;

each R¹ is independently selected from hydrogen, halo and haloalkyl;

R⁴ is hydrogen, C₁₋₈ alkyl, and aryl, substituted with n occurrences ofR²;

each R² is independently selected from halo, haloalkyl, alkyl, aryl,heteroaryl, aralkyl, heteroaralkyl, cyano, —OR^(a), —COOR^(b) and—CONR^(c)R^(c′); wherein two R² together with the carbons to which theyare attached, may form an optionally substituted ring, each of which canbe further substituted;

each R³ is independently selected from halo, haloalkyl and —OR^(a);

R⁵ is hydrogen or C₁₋₈ alkyl;

R^(a) is independently selected from alkyl, haloalkyl, optionallysubstituted heteroaryl and optionally substituted heterocyclyl;

each R^(b) is independently alkyl;

each R^(c) is independently selected from hydrogen and alkyl; and

n is 0, 1, 2, or 3.

In some embodiments, B, D and E are each independently H.

In some embodiments, each R¹ is independently H. In some embodiments,each R¹ is independently halo (e.g., chloro). In some embodiments, eachR¹ is independently haloalkyl (e.g., trifluoromethyl).

In some embodiments, one R¹ is halo and one R¹ is haloalkyl. In someembodiments, one R¹ is chloro and one R¹ is trifluoromethyl.

In some embodiments, R⁵ is hydrogen.

In some embodiments, R⁵ is C₁₋₈ alkyl (e.g., methyl).

In some embodiments, R⁴ is hydrogen.

In some embodiments, R⁴ is C₁₋₈ alkyl or aryl substituted with noccurrences of R². In some embodiments, R⁴ is C₁₋₈ alkyl (e.g., methylor ethyl) substituted with n occurrences of R².

In some embodiments, each R² is independently selected from halo,haloalkyl, alkyl, aryl, heteroaryl, heteroaralkyl, cyano, —OR^(a),—COOR^(b) and —CONR^(c)R^(c′); wherein two R², together with the carbonsto which they are attached, may form an optionally substituted ring,each of which can be further substituted.

In some embodiments, n is 0. In some embodiments, n is 1. In someembodiments, R² is —OR^(a). In some embodiments, R^(a) is alkyl (e.g.,methyl). In some embodiments, R² is optionally substituted heteroaryl.In some embodiments, R² is optionally substituted monocyclic heteroaryl(e.g., 3-pyridyl). In some embodiments, R² is optionally substitutedaryl. In some embodiments, R² is optionally substituted monocyclic aryl(e.g., 4-chlorophenyl).

In some embodiments, R⁴ is aryl (e.g., phenyl) substituted with noccurrences of R². In some embodiments, R⁴ is phenyl substituted with noccurrences of R². In some embodiments, n is 0.

In some embodiments, n is 1. In some embodiments, R² is halo (e.g.,fluoro or chloro). In some embodiments, R² is haloalkyl (e.g.,trifluoromethyl). In some embodiments, R² is alkyl (e.g., methyl orethyl). In some embodiments, R² is heteoaralkyl. In some embodiments, R²is optionally substituted monocyclic heteroaralkyl (e.g.,methyl-4-trifluoromethyl-2-pyridyl). In some embodiments, R² is cyano.In some embodiments, R² is —OR^(a). In some embodiments, R^(a) is alkyl(e.g., methyl). In some embodiments, R² is —COOR^(b). In someembodiments, R^(b) is alkyl (e.g., ethyl). In some embodiments, R² isoptionally substituted monocyclic heteroaryl. In some embodiments, R² isoptionally substituted pyridyl. In some embodiments, R² is pyridylsubstituted with haloalkyl (e.g., trifluoromethyl).

In some embodiments, n is 2. In some embodiments, both R² are halo(e.g., fluoro or chloro). In some embodiments, n is 2. In someembodiments, both R² are alkyl (e.g., methyl). In some embodiments, n is2. In some embodiments, one R² is halo (e.g., fluoro or chloro) and oneis alkyl (e.g., methyl). In some embodiments, n is 2. In someembodiments, one R² is halo and one is —CONR^(c)R^(c′). In someembodiments, n is 2. In some embodiments, one R² is chloro and one is—CONHR^(c′). In some embodiments, R^(c) is alkyl (e.g., methyl orisopropyl). In some embodiments, n is 2. In some embodiments, one R² isalkyl and one is —CONR^(c)R^(c′). In some embodiments, n is 2. In someembodiments, one R² is methyl and one is —CONHR^(c′). In someembodiments, R^(c) is alkyl (e.g., methyl or isopropyl).

In some embodiments, n is 2. In some embodiments, one R² is haloalkyl(e.g., trifluoromethyl) and the other is —OR^(a). In some embodiments,R^(a) is alkyl (e.g., methyl). In some embodiments, n is 2. In someembodiments, one R² is halo and the other is —OR^(a). In someembodiments, n is 2. In some embodiments, one R² is chloro and the otheris —OR^(a). In some embodiments, R^(a) is optionally substitutedheteroaryl. In some embodiments, n is 2. In some embodiments, both R²are —OR^(a). In some embodiments, R^(a) is alkyl (e.g., methyl). In someembodiments, R^(a) is optionally substituted pyridyl. In someembodiments, R^(a) is pyridyl substituted with haloalkyl (e.g.,trifluoromethyl). In some embodiments, R^(a) is optionally substitutedheterocyclyl. In some embodiments, R^(a) is an optionally substituted5-membered heterocyclyl. In some embodiments, R^(a) is optionallysubstituted pyrrolidinyl. In some embodiments, R^(a) is N-methylpyrrolidinyl. In some embodiments, R^(a) is:

In some embodiments, n is 2. In some embodiments, two R², together withthe carbon atoms to which they are attached, form a 5-memberedheterocyclic ring. In some embodiments, two R², together with the phenylring to which they are attached, form the following structure:

In some embodiments, n is 3. In some embodiments, each R² is halo (e.g.,fluoro). In some embodiments, n is 3. In some embodiments, two R² arehalo and one R² is —CONR^(c)R^(c′). In some embodiments, two R² arechloro and one R² is —CONHR^(c). In some embodiments, R^(c) is alkyl(e.g., methyl or isopropyl). In some embodiments, one R² is chloro, oneR² is bromo, and one R² is —CONHR^(c′). In some embodiments, R^(c) isalkyl (e.g., methyl or isopropyl). In some embodiments, n is 3. In someembodiments, one R² is halo, one R² is alkyl, and one R² is—CONR^(c)R^(c′). In some embodiments, one R² is chloro, one R² ismethyl, and one R² is —CONHR^(c′). In some embodiments, R^(c) is alkyl(e.g., methyl or isopropyl). In some embodiments, one R² is bromo, oneR² is methyl, and one R² is —CONHR^(c′). In some embodiments, R^(c) isalkyl (e.g., methyl or isopropyl).

In some embodiments, R³ is halo (e.g., chloro or bromo). In someembodiments, R³ is haloalkyl (e.g., trifluoromethyl). In someembodiments, R³ is —OR^(a). In some embodiments, R^(a) is haloalkyl(e.g., difluoromethoxy, trifluoromethoxy or trifluoroethoxy). In someembodiments, R^(a) is —CH₂CF₃.

In another aspect, the invention features a pharmaceutical compositioncomprising a compound of formula (Va):

wherein

B and D are each independently selected from H and SO₂NR⁴R⁵; wherein atleast one of B or D is —SO₂—NR⁴R⁵;

A and E are each independently selected from H and R³;

each R¹ is independently selected from hydrogen, halo and haloalkyl;

R⁴ is hydrogen, C₁₋₈ alkyl, and aryl, substituted with n occurrences ofR²;

each R² is independently selected from halo, haloalkyl, alkyl, aryl,heteroaryl, aralkyl, heteroaralkyl, cyano, —OR^(a), —COOR^(b) and—CONR^(c)R^(c′); wherein two R², together with the carbons to which theyare attached, may form an optionally substituted ring, each of which canbe further substituted;

each R³ is independently selected from halo, haloalkyl and —OR^(a);

R⁵ is hydrogen or C₁₋₈ alkyl;

R^(a) is independently selected from alkyl, haloalkyl, optionallysubstituted heteroaryl and optionally substituted heterocyclyl;

each R^(b) is independently alkyl;

each R^(c) is independently selected from hydrogen and alkyl; and

n is 0, 1, 2, or 3.

In another aspect, the invention features a method of treating adisorder described herein (e.g., cancer) comprising administering to asubject a compound of formula (V) or (Va) as described herein or apharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of preventing (e.g.,preventing the onset of at least one symptom) or delaying the onset of adisorder described here (e.g., cancer) comprising administering to asubject a compound of formula (V) or (Va) as described herein or apharmaceutically acceptable salt thereof.

In some embodiments, A and E are each H.

In some embodiments, B is SO₂NR⁴R⁵ and D is H. In some embodiments, B isH and D is SO₂NR⁴R⁵.

In one aspect, the present invention features a compound orpharmaceutically acceptable salt thereof of formula (VI):

wherein

A, B and E are each independently selected from H, —SO₂—NR⁴R⁵ and R³;wherein at least one of A, B or E is —SO₂—NR⁴R⁵;

Y¹, Y², Y³ and Y⁴ are each independently selected from N and CR¹,wherein at least one of Y¹, Y², Y³ and Y⁴ are N;

each R⁴ is independently selected from C₁₋₈ alkyl, aryl and heteroaryl,each of which is substituted with n occurrences of R²;

each R⁵ is independently hydrogen or C₁₋₈ alkyl;

each R¹ is independently selected from hydrogen, C₁₋₈ alkyl, C₁₋₈terminal alkynyl, C₁₋₈ alkoxy, halogen, haloalkyl and haloalkoxy;

each R² is independently selected from halo, haloalkyl, C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alknynyl, aryl, heteroaryl, aralkyl, heteroaralkyl, cyano,—OR^(a), —COOR^(b) and —CONR^(c)R^(c′); wherein two R², together withthe carbons to which they are attached, may form an optionallysubstituted ring, each of which can be further substituted;

each R³ is independently selected from C₁₋₈ alkyl, —OR^(a), halogen,haloalkyl, haloalkoxy and optionally substituted heteroaryl;

each R^(a) is independently selected from alkyl, haloalkyl, optionallysubstituted heteroaryl and optionally substituted heterocyclyl;

each R^(b) is independently alkyl; and

each R^(c) is independently selected from hydrogen and alkyl; and

n is 0, 1, 2 or 3.

In some embodiments, at least one of Y¹, Y², Y³ and Y⁴ is N. In someembodiments, at least one of Y¹, Y², Y³ and Y⁴ are CH. In someembodiments, Y¹ is N. In some embodiments, Y³ is N.

In some embodiments, each R¹ is independently hydrogen.

In some embodiments, the invention features a compound of formula (VIa):

wherein n, B, E, R¹, R², R³, R⁴ and R⁵ are defined as above.

In some embodiments, the invention features a compound of formula (VIb):

wherein n, B, E, R¹, R², R³, R⁴ and R⁵ are defined as above.

In some embodiments, B and E are each independently selected from H.

In some embodiments, R⁵ is hydrogen.

In some embodiments, each R¹ is independently hydrogen. In someembodiments, each R¹ is independently selected from C₁₋₈ alkyl, halogenor haloalkyl. In some embodiments, each R¹ is independently selectedfrom halogen or haloalkyl. In some embodiments, each R¹ is independentlyselected from halogen (e.g., chlorine or fluorine). In some embodiments,each R¹ is independently haloalkyl (e.g., trifluoroalkyl).

In some embodiments, R⁴ is selected from aryl or heteroaryl. In someembodiments, R⁴ is aryl substituted with n occurrences of R². In someembodiments, R⁴ is C₅₋₈ monocyclic aryl or C₈₋₁₄ bicyclic aryl. In someembodiments, R⁴ is C₅₋₈ monocyclic aryl (e.g., optionally substitutedphenyl). In some embodiments, R⁴ is phenyl substituted with noccurrences of R². In some embodiments, R² is heteroaryl substitutedwith n occurrences of R². In some embodiments, R⁴ is a 5-8 memberedheteroaryl or 8-14 membered heteroaryl. In some embodiments, R⁴ is an8-12 membered heteroaryl (e.g., 5-quinolyl or 6-quinolyl). In someembodiments, R⁴ is quinolyl (e.g., 5-quinolyl or 6-quinolyl) substitutedwith n occurrences of R².

In some embodiments, n is 0. In some embodiments, n is 1.

In some embodiments, R² is halo, C₁₋₄ alkyl or haloalkyl, each of whichcan be further substituted.

In some embodiments, R² is C₁₋₄ alkyl (e.g., ethyl). In someembodiments, R² is halo (e.g., fluoro or chloro). In some embodiments,R² is haloalkyl (e.g., trifluoromethyl).

In some embodiments, n is 2. In some embodiments, both R² are C₁₋₄ alkyl(e.g., methyl). In some embodiments, n is 2. In some embodiments, bothR² are halo (e.g., fluoro or chloro). In some embodiments, n is 2. Insome embodiments, one R² is haloalkyl (e.g., trifluoroalkyl) and theother R² is —OR^(a). In some embodiments, R^(a) is alkyl (e.g., methylor ethyl). In some embodiments, n is 2. In some embodiments, on R² ishalo (e.g., fluoro or chloro) and the other R² is C₁₋₄ alkyl (e.g.,methyl or ethyl).

In some embodiments, n is 2. In some embodiments, two R², together withthe carbon atoms to which they are attached, form a 5-memberedheterocyclic ring. In some embodiments, two R², together with the phenylring to which they are attached, form the following structure:

In some embodiments, n is 3. In some embodiments, all R² are halo (e.g.,fluoro or chloro).

In another aspect, the invention features a pharmaceutical compositioncomprising a compound selected from Formula (VI), (VIa) or (VIb) asdescribed herein or a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of treating adisorder described herein (e.g., cancer) comprising administering to asubject a compound of formula (VI), (VIa) or (VIb) as described hereinor a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of preventing (e.g.,preventing the onset of at least one symptom) or delaying the onset of adisorder as described herein (e.g., cancer) comprising administering toa subject a compound of formula (VI), (VIa) or (VIb) as described hereinor a pharmaceutically acceptable salt thereof.

In one aspect, the present invention features a compound orpharmaceutically acceptable salt thereof of formula (VII):

wherein

-   -   X¹ is N or CE;    -   X² is N or CD;    -   X³ is N or CB;    -   X⁴ is N or CA, wherein at least one of X¹, X², X³ and X⁴ is N        and at least one of X¹, X², X³, X⁴, is C—SO₂—NR⁴R⁵;    -   A, B, D and E are each independently selected from H, R³ and        —SO₂—NR⁴R⁵;    -   Y¹ is selected from N and CR¹;    -   each R⁴ is independently selected from C₁₋₈ alkyl, aryl and        heteroaryl, each of which is substituted with n occurrences of        R²;    -   R⁵ is hydrogen or C₁₋₈ alkyl;    -   each R¹ is independently selected from hydrogen, C₁₋₈ alkyl,        C₁₋₈ terminal alkynyl, C₁₋₈ alkoxy, halogen, haloalkyl and        haloalkoxy;    -   each R² is independently selected from halo, haloalkyl, C₁₋₄        alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, aryl, heteroaryl, aralkyl,        heteroaralkyl, cyano, —OR^(a), —COOR^(b) and —CONR^(c)R^(c′);        wherein two R², together with the carbons to which they are        attached, may form an optionally substituted ring, each of which        can be further substituted;    -   each R³ is independently selected from C₁₋₈ alkyl, —OR^(a),        halogen, haloalkyl, haloalkoxy and optionally substituted        heteroaryl;    -   each R^(a) is independently selected from alkyl, haloalkyl,        optionally substituted heteroaryl and optionally substituted        heterocyclyl;    -   each R^(b) is independently alkyl; and    -   each R^(c) is independently selected from hydrogen and alkyl;        and    -   n is 0, 1, 2 or 3.

In some embodiments, the invention features a compound of formula(VIIa):

wherein n, B, D, R¹, R⁴, R² and R⁵ are defined as in formula (VII).

In some embodiments, the invention features a compound of formula(VIIa):

wherein n, B, E, R¹, R⁴, R² and R⁵ are defined as in formula (VII).

In some embodiments, the invention features a compound of formula(VIIb):

wherein n, B, D, R¹, R⁴, R² and R⁵ are defined as in formula (VII).

In some embodiments, the invention features a compound of formula(VIIa):

wherein n, B, E, R¹, R⁴, R² and R⁵ are defined as in formula (VII).

In some embodiments, B and E are each independently selected from H.

In some embodiments, B and D are each independently selected from H.

In some embodiments, R⁵ is hydrogen.

In some embodiments, each R¹ is independently hydrogen. In someembodiments, each R¹ is independently selected from C₁₋₈ alkyl, halogenor haloalkyl. In some embodiments, each R¹ is independently selectedfrom halogen or haloalkyl. In some embodiments, each R¹ is independentlyselected from halogen (e.g., chlorine or fluorine). In some embodiments,each R¹ is independently haloalkyl (e.g., trifluoroalkyl).

In some embodiments, R⁴ is selected from aryl or heteroaryl. In someembodiments, R⁴ is aryl substituted with n occurrences of R². In someembodiments, R⁴ is C₅₋₈ monocyclic aryl or C₈₋₁₄ bicyclic aryl. In someembodiments, R⁴ is C₅₋₈ monocyclic aryl (e.g., optionally substitutedphenyl). In some embodiments, R⁴ is phenyl substituted with noccurrences of R².

In some embodiments, n is 0. In some embodiments, n is 1.

In some embodiments, R² is halo, C₁₋₄ alkyl or haloalkyl, each of whichcan be further substituted.

In some embodiments, R² is C₁₋₄ alkyl (e.g., methyl or ethyl). In someembodiments, R² is halo (e.g., fluoro or chloro).

In some embodiments, n is 2. In some embodiments, both R² are C₁₋₄ alkyl(e.g., methyl or ethyl). In some embodiments, n is 2. In someembodiments, both R² are halo (e.g., fluoro or chloro). In someembodiments, n is 2. In some embodiments, one R² is haloalkyl (e.g.,trifluoroalkyl) and the other R² is —OR^(a). In some embodiments, R^(a)is alkyl (e.g., methyl or ethyl). In some embodiments, n is 2. In someembodiments, one R² is C₁₋₄ alkyl (e.g., methyl or ethyl) and the otherR² is halo (e.g., fluoro or chloro).

In some embodiments, n is 2. In some embodiments, two R², together withthe carbon atoms to which they are attached, form a 5-memberedheterocyclic ring. In some embodiments, two R², together with the phenylring to which they are attached, form the following structure:

In some embodiments, n is 3. In some embodiments, all R² are halo (e.g.,fluoro or chloro).

In another aspect, the invention features a pharmaceutical compositioncomprising a compound selected from formula (VII), (VIIa), (VIIb),(VIIc) or (VIId) as described herein or a pharmaceutically acceptablesalt thereof.

In another aspect, the invention features a method of treating adisorder described herein (e.g., cancer) comprising administering to asubject a compound of formula (VII), (VIIa), (VIIb), (VIIc) or (VIId) asdescribed herein or a pharmaceutically acceptable salt thereof.

In another aspect, the invention features a method of preventing (e.g.,preventing the onset of at least one symptom) or delaying the onset of adisorder described here (e.g., cancer) comprising administering to asubject a compound of formula (VII), (VIIa), (VIIb), (VIIc) or (VIId) asdescribed herein or a pharmaceutically acceptable salt thereof.

In one aspect, the invention features a method of modulating (e.g.,increasing or decreasing) the level of PKM2 activity and/or glycolysis(e.g., modulating the endogenous ability of a cell in the patient todown regulate PKM2) in a patient in need thereof. The method comprisesthe step of administering an effective amount of a compound describedherein to the patient in need thereof, thereby modulating (e.g.,increasing or decreasing) the level of PKM2 activity and/or glycolysisin the patient. In some embodiments of the invention an activator isused to maintain PKM2 in its active conformation or activate pyruvatekinase activity in proliferating cells as a means to divert glucosemetabolites into catabolic rather than anabolic processes in thepatient.

In another aspect, the invention features a method of regulating cellproliferation in a patient in need thereof. The method comprises thestep of administering an effective amount of a compound described hereinto the patient in need thereof, thereby regulating cell proliferation inthe patient. E.g., this method can modulate growth of a transformedcell, e.g., a cancer cell, or generally modulate growth in aPKM2-dependent cell that undergoes aerobic glycolysis.

In another aspect, the invention features a method of treating a patientsuffering from or susceptible to a disease or disorder associated withthe function of PKM2 in a patient in need thereof. The method comprisesthe step of administering an effective amount of a compound describedherein to the patient in need thereof, thereby treating, preventing orameliorating the disease or disorder in the patient. In anotherembodiment the modulator is provided in a pharmaceutical composition.

In another embodiment the method includes identifying or selecting apatient who would benefit from modulation (e.g., activation orinhibition) of PKM2. E.g., the patient can be identified on the basis ofthe level of PKM2 activity in a cell of the patient (e.g., as opposed tomerely being in need of treatment of the disorder itself, e.g., cancer).In another embodiment the selected patient is a patient suffering fromor susceptible to a disorder or disease identified herein, e.g., adisorder characterized by unwanted cell growth or proliferation, e.g.,cancer, obesity, diabetes, atherosclerosis, restenosis, and autoimmunediseases.

In one aspect, the invention features a method of evaluating a candidatecompound the method comprising:

optionally supplying the candidate compound;

contacting the compound with a cell, e.g., a cell having an intactplasma membrane;

evaluating the ability of the compound to interact intracellularly with,e.g., to form a complex with, to bind, e.g., specifically to, or tomodulate (e.g., activate or inhibit) the activity of, a target kinase,e.g., pyruvate kinase, e.g., PKM2;

thereby evaluating the candidate compound.

In some embodiments, evaluating the candidate compound comprisesevaluating the candidate compound for use as an anti-proliferative oranti-cancer agent and the ability of the candidate compound to interactintracellulary with the target compound is correlated to efficacy as ananti-proliferative or anti-cancer agent.

In some embodiments, evaluating the candidate compound comprisesevaluating the ability of the candidate compound cross the cell membraneand the ability of the candidate compound to interact intracellularywith the target compound is correlated to with ability of the candidatecompound to cross the cell membrane.

In some embodiments, evaluating the candidate compound comprisesevaluating the ability of the candidate compound to modulate any of thefollowing properties: a conformational state in the target kinase,binding of the target kinase to an endogenous modulator of target kinaseactivity, e.g., FBP or a phosphotyrosine containing polypeptide, orother property of a target kinase disclosed herein, and the ability ofthe candidate compound to interact with the target compound iscorrelated with one or more of said properties.

In some embodiments, the method further comprises separating the cellfrom candidate compound that has not entered the cell, e.g., by washingthe cell or removing the cell from an animal to which the candidatecompound has been administered.

In some embodiments, the method further comprises lysing (e.g., bydisrupting or dissolving the cell membrane) the cell, e.g., prior toevaluating the ability of the candidate compound to interact with thetarget kinase.

In some embodiments, contacting the candidate compound with a cellcomprises contacting the candidate compound with a whole animal, atissue that is not part of a whole animal, an organ which is not part ofa whole animal or a cell which is not part of a whole animal.

In some embodiments, the cell is a cultured cell, e.g., a primary cell,a secondary cell.

In some embodiments, the cell is a mammal, primate, human, rodent,mouse, rat, or hamster cell.

In some embodiments, the cell is a tumor or transformed cell, e.g., asolid tumor cell.

In some embodiments, a plurality of target compounds are evaluated,e.g., at least 10, 20, 50, 100, or 500 candidate compounds areevaluated.

In some embodiments, a plurality of candidate compounds are evaluatedsimultaneously, e.g., wherein each of a plurality of candidate compoundsis evaluated individually but simultaneously, or wherein a plurality ofcandidate compounds are pooled and contacted with the same cell or samealiquot of cells.

In some embodiments, a plurality of candidate compounds are evaluated inan automated device.

In some embodiments, evaluating a candidate compound comprises providinga value for the ability of the candidate compound to interact with thetarget kinase and, optionally, comparing that value to a predeterminedvalue, e.g., a value for a positive and or negative control.

In some embodiments, the method further comprises selecting a candidatecompound have a value for interacting with the target kinase which has apreselected relationship with a reference value, e.g., the value for thecandidate compound exceeds a preselected minimum value, e.g., apreselected minimum value for activation of the target kinase.

In some embodiments, the method further comprises evaluating, e.g.,confirming, the ability of a candidate compound (e.g., a candidatecompound which meets a predetermined level of interaction (e.g., complexformation, specific binding, or modulation (e.g., activation orinhibition) in the evaluating step) to interact with, e.g., to form acomplex with, to bind specifically to, or to modulate (e.g., activate orinhibit) the activity of the target kinase in a second method.

In some embodiments, the method further comprises selecting a candidatecompound and repeating the evaluation under the same or differentconditions, e.g., at the same or a different concentration.

In some embodiments, the method further comprises selecting a candidatecompound evaluated in a cell other than a whole animal and confirmingthe activity determined in the cell-based assay by evaluation in a wholeanimal.

In some embodiments, the method further comprises selecting a candidatecompound and confirming the activity determined in the evaluation by asecond, different assay.

In some embodiments, a plurality of structurally related candidatecandidates are evaluated, e.g., a plurality of candidate candidateshaving a common core or scaffold.

In some embodiments, the method comprises providing a plurality ofsecond generation candidate candidates which are analogs of a candidatecompound.

In some embodiments, the method comprises evaluating a first candidatecompound, comparing the structure of the first candidate compound to asecond candidate compound and evaluating the second candidate compoundIn some embodiments, the candidate compound is contacted with culturedcells, e.g., cultured cells having a preselected level of confluency,e.g., from about 60% to about 95%, preferably from about 70% to about90% confluent.

In some embodiments, the candidate compound is contacted with the cellfor a preselected length of time, e.g., a time period sufficient toallow a positive control to enter the cell and interact with the targetkinase.

In some embodiments, the contacting step comprises contacting thecompound with the cell for at least about 0.1, 0.5, 1, 2, 3, 4, 5, or 6hours.

In some embodiments, the compound forms a complex with the targetkinase.

In some embodiments, the compound binds, e.g., specifically to thetarget kinase.

In some embodiments, the target kinase is PKM2 and the candidatecompound induces a conformational change (e.g., from a non-activated orless activated conformation to an activated or more activatedconformation or from an activated or more activated conformation to anon-activated or less activated conformation) in the target kinase.

In some embodiments, the target kinase is PKM2 and the candidatecompound increases the activity of PKM2.

In some embodiments, the target kinase is PKM2 and the candidatecompound decreases the activity of PKM2.

In some embodiments, the method further comprises evaluating thepresence and/or amount of lactate, e.g., in the media.

In some embodiments, the lysing step comprises snap-freezing the cell,e.g., on dry ice.

In some embodiments, the lysing step comprises adding a lysis buffer,e.g., a detergent-containing (e.g., Triton-containing) lysis buffer(e.g., a lysis buffer described in Table 3), to the cell.

In some embodiments, the detergent is used at a concentration that doesnot disrupt the interaction (e.g., binding) between the compound and thekinase, e.g., pyruvate kinase, e.g., PKM2, e.g., at no more than about0.1, 0.5, 1, 1.5, 2, or 5%.

In one aspect, the invention features a method of evaluating a candidatecompound the method comprising:

optionally supplying the candidate compound;

contacting the candidate compound with a cell, which cell is outside ananimal, e.g., a cell having an intact plasma membrane;

separating the cell from candidate compound that has not entered thecell;

lysing said cell under conditions that do not abolish the binding of thecandidate compound to PKM2; and

evaluating the ability of the compound to interact intracellularly with,e.g., to form a complex with, to bind, e.g., specifically to, or tomodulate (e.g., activate or inhibit) the activity of PKM2;

thereby evaluating the candidate compound.

In one aspect, the invention features a method of evaluating a candidatecompound the method comprising:

optionally supplying the candidate compound;

contacting the candidate compound with a cell which is part of a wholeanimal;

lysing said cell under conditions that do not abolish the binding of thecandidate compound to PKM2; and

evaluating the ability of the compound to interact intracellularly with,e.g., to form a complex with, to bind, e.g., specifically to, or tomodulate (e.g., activate or inhibit) the activity of PKM2;

thereby evaluating the candidate compound.

In another embodiment the compound described herein is administered at adosage and frequency sufficient to increase lactate production oroxidative phosphorylation.

The term “halo” or “halogen” refers to any radical of fluorine,chlorine, bromine or iodine.

The term “alkyl” refers to a hydrocarbon chain that may be a straightchain or branched chain, containing the indicated number of carbonatoms. For example, C₁-C₁₂ alkyl indicates that the group may have from1 to 12 (inclusive) carbon atoms in it. The term “haloalkyl” refers toan alkyl in which one or more hydrogen atoms are replaced by halo, andincludes alkyl moieties in which all hydrogens have been replaced byhalo (e.g., perfluoroalkyl). The terms “arylalkyl” or “aralkyl” refer toan alkyl moiety in which an alkyl hydrogen atom is replaced by an arylgroup. Aralkyl includes groups in which more than one hydrogen atom hasbeen replaced by an aryl group. Examples of “arylalkyl” or “aralkyl”include benzyl, 2-phenylethyl, 3-phenylpropyl, 9-fluorenyl, benzhydryl,and trityl groups.

The term “alkylene” refers to a divalent alkyl, e.g., —CH₂—, —CH₂CH₂—,and —CH₂CH₂CH₂—.

The term “alkenyl” refers to a straight or branched hydrocarbon chaincontaining 2-12 carbon atoms and having one or more double bonds.Examples of alkenyl groups include, but are not limited to, allyl,propenyl, 2-butenyl, 3-hexenyl and 3-octenyl groups. One of the doublebond carbons may optionally be the point of attachment of the alkenylsubstituent. The term “alkynyl” refers to a straight or branchedhydrocarbon chain containing 2-12 carbon atoms and characterized inhaving one or more triple bonds. Examples of alkynyl groups include, butare not limited to, ethynyl, propargyl, and 3-hexynyl. One of the triplebond carbons may optionally be the point of attachment of the alkynylsubstituent.

The terms “alkylamino” and “dialkylamino” refer to —NH(alkyl) and—NH(alkyl)₂ radicals respectively. The term “aralkylamino” refers to a—NH(aralkyl) radical. The term alkylaminoalkyl refers to a(alkyl)NH-alkyl- radical; the term dialkylaminoalkyl refers to a(alkyl)₂N-alkyl- radical The term “alkoxy” refers to an —O-alkylradical. The term “mercapto” refers to an SH radical. The term“thioalkoxy” refers to an —S-alkyl radical. The term thioaryloxy refersto an —S-aryl radical.

The term “aryl” refers to an aromatic monocyclic, bicyclic, or tricyclichydrocarbon ring system, wherein any ring atom capable of substitutioncan be substituted (e.g., by one or more substituents). Examples of arylmoieties include, but are not limited to, phenyl, naphthyl, andanthracenyl.

The term “cycloalkyl” as employed herein includes saturated cyclic,bicyclic, tricyclic, or polycyclic hydrocarbon groups having 3 to 12carbons. Any ring atom can be substituted (e.g., by one or moresubstituents). The cycloalkyl groups can contain fused rings. Fusedrings are rings that share a common carbon atom. Examples of cycloalkylmoieties include, but are not limited to, cyclopropyl, cyclohexyl,methylcyclohexyl, adamantyl, and norbornyl.

The term “heterocyclyl” refers to a nonaromatic 3-10 memberedmonocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ringsystem having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms ifbicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selectedfrom O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms ofN, O, or S if monocyclic, bicyclic, or tricyclic, respectively). Theheteroatom may optionally be the point of attachment of the heterocyclylsubstituent. Any ring atom can be substituted (e.g., by one or moresubstituents). The heterocyclyl groups can contain fused rings. Fusedrings are rings that share a common carbon atom. Examples ofheterocyclyl include, but are not limited to, tetrahydrofuranyl,tetrahydropyranyl, piperidinyl, morpholino, pyrrolinyl, pyrimidinyl,quinolinyl, and pyrrolidinyl.

The term “cycloalkenyl” refers to partially unsaturated, nonaromatic,cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon groups having 5to 12 carbons, preferably 5 to 8 carbons. The unsaturated carbon mayoptionally be the point of attachment of the cycloalkenyl substituent.Any ring atom can be substituted (e.g., by one or more substituents).The cycloalkenyl groups can contain fused rings. Fused rings are ringsthat share a common carbon atom. Examples of cycloalkenyl moietiesinclude, but are not limited to, cyclohexenyl, cyclohexadienyl, ornorbornenyl.

The term “heterocycloalkenyl” refers to a partially saturated,nonaromatic 5-10 membered monocyclic, 8-12 membered bicyclic, or 11-14membered tricyclic ring system having 1-3 heteroatoms if monocyclic, 1-6heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, saidheteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6,or 1-9 heteroatoms of N, O, or S if monocyclic, bicyclic, or tricyclic,respectively). The unsaturated carbon or the heteroatom may optionallybe the point of attachment of the heterocycloalkenyl substituent. Anyring atom can be substituted (e.g., by one or more substituents). Theheterocycloalkenyl groups can contain fused rings. Fused rings are ringsthat share a common carbon atom. Examples of heterocycloalkenyl includebut are not limited to tetrahydropyridyl and dihydropyranyl.

The term “heteroaryl” refers to an aromatic 5-8 membered monocyclic,8-12 membered bicyclic, or 11-14 membered tricyclic ring system having1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9heteroatoms if tricyclic, said heteroatoms selected from O, N, or S(e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S ifmonocyclic, bicyclic, or tricyclic, respectively). Any ring atom can besubstituted (e.g., by one or more substituents).

The term “oxo” refers to an oxygen atom, which forms a carbonyl whenattached to carbon, an N-oxide when attached to nitrogen, and asulfoxide or sulfone when attached to sulfur.

The term “acyl” refers to an alkylcarbonyl, cycloalkylcarbonyl,arylcarbonyl, heterocyclylcarbonyl, or heteroarylcarbonyl substituent,any of which may be further substituted (e.g., by one or moresubstituents).

The term “substituents” refers to a group “substituted” on an alkyl,cycloalkyl, alkenyl, alkynyl, heterocyclyl, heterocycloalkenyl,cycloalkenyl, aryl, or heteroaryl group at any atom of that group. Anyatom can be substituted. Suitable substituents include, withoutlimitation, alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11,C12 straight or branched chain alkyl), cycloalkyl, haloalkyl (e.g.,perfluoroalkyl such as CF₃), aryl, heteroaryl, aralkyl, heteroaralkyl,heterocyclyl, alkenyl, alkynyl, cycloalkenyl, heterocycloalkenyl,alkoxy, haloalkoxy (e.g., perfluoroalkoxy such as OCF₃), halo, hydroxy,carboxy, carboxylate, cyano, nitro, amino, alkyl amino, SO₃H, sulfate,phosphate, methylenedioxy (—O—CH₂—O— wherein oxygens are attached tovicinal atoms), ethylenedioxy, oxo, thioxo (e.g., C═S), imino (alkyl,aryl, aralkyl), S(O)_(n)alkyl (where n is 0-2), S(O)_(n) aryl (where nis 0-2), S(O)_(n) heteroaryl (where n is 0-2), S(O)_(n) heterocyclyl(where n is 0-2), amine (mono-, di-, alkyl, cycloalkyl, aralkyl,heteroaralkyl, aryl, heteroaryl, and combinations thereof), ester(alkyl, aralkyl, heteroaralkyl, aryl, heteroaryl), amide (mono-, di-,alkyl, aralkyl, heteroaralkyl, aryl, heteroaryl, and combinationsthereof), sulfonamide (mono-, di-, alkyl, aralkyl, heteroaralkyl, andcombinations thereof). In one aspect, the substituents on a group areindependently any one single, or any subset of the aforementionedsubstituents. In another aspect, a substituent may itself be substitutedwith any one of the above substituents.

The term “selective” is meant at least 2-fold, 3-fold, 4-fold, 5-fold,6-fold, or 10-fold greater modulation (e.g., inhibition) of M2 than M1.

The term “activator” as used herein means an agent that (measurably)increases the activity of a pyruvate kinase (e.g., PKM2) or causespyruvate kinase (e.g., PKM2) activity to increase to a level that isgreater than PKM2's basal levels of activity. For example, the activatormay mimic the effect caused by a natural ligand (e.g., FBP). Theactivator effect caused by the agent may be to the same, or to agreater, or to a lesser extent than the activating effect caused by anatural ligand, but the same type of effect is caused. Peptides, nucleicacids, and small molecules may be activators. An agent can be evaluatedto determine if it is an activator by measuring either directly orindirectly the activity of the pyruvate kinase when subjected to theagent. The activity of the agent can be measured, for example, against acontrol substance. In some instances, the activity measured of the agentis for activation of PKM2. The activity of PKM2 can be measured, forexample, by monitoring the concentration of a substrate such as ATP orNADH.

The term “inhibitor” as used herein means an agent that measurablyslows, stops, decreases or inactivates the enzymatic activity ofpyruvate kinase (e.g., PKM2) to decrease to a level that is less thanthe pyruvate kinase's (e.g., PKM2's) basal levels of activity.Inhibitors of pyruvate kinase (e.g., PKM2) may be peptides or nucleicacids. An agent can be evaluated to determine if it is an inhibitor bymeasuring either directly or indirectly the activity of the pyruvatekinase when subjected to the agent. The activity of the agent can bemeasured, for example, against a control substance. In some instances,the activity measured of the agent is for inhibition of PKM2. Theactivity of PKM2 can be measured, for example, by monitoring theconcentration of a substrate such as ATP or NADH.

The term “modulate” refers to an increase or decrease, e.g., in theactivity of an enzyme in response to exposure to a compound orcomposition described herein, e.g., the activation or inhibition ofPKM2, in at least a sub-population of cells in a subject such that adesired end result is achieved (e.g., a therapeutic result). In someembodiments, a compound as described herein inhibits a target describedherein, e.g., PKM2. In some embodiments, a compound as described hereinis activates a target described herein, e.g., PKM2.

DETAILED DESCRIPTION

This invention is not limited in its application to the details ofconstruction and the arrangement of components set forth in thefollowing description or illustrated in the drawings. The invention iscapable of other embodiments and of being practiced or of being carriedout in various ways. Also, the phraseology and terminology used hereinis for the purpose of description and should not be regarded aslimiting. The use of “including,” “comprising,” or “having,”“containing”, “involving”, and variations thereof herein, is meant toencompass the items listed thereafter and equivalents thereof as well asadditional items.

Compounds

Described herein are compounds and compositions that modulate PKM2, forexample, activate or inhibit PKM2. Compounds that modulate PKM2, e.g.,activate or inhibit PKM2, can be used to treat disorders such asneoplastic disorders (e.g., cancer) or fat related disorders (e.g.,obesity). Exemplary compounds include the compounds of Formulas (I),(II), (III), (IV), (V), (VI) and (VII) described herein. In someembodiments, a compound described herein modulates PKM2 by interacting(e.g., binding) with the FBP binding pocket. For example, a compounddescribed herein can compete with FBP binding in PKM2.

In some embodiments a compound described herein has one or moreproperties described herein, e.g., one or more of the followingproperties: it is an allosteric modulator (e.g., inhibitor oractivator); it modulates the release of FBP (e.g., inhibits orpromotes); it is a modulator (e.g., agonist or antagonist) of FBP, e.g.,an agonist which binds with a lower, about the same, or higher affinitythan does FBP; it modulates (e.g., inhibits or promotes) the dissolutionof tetrameric PKM2; it modulates (e.g., promotes or inhibits) theassembly of tetrameric PKM2; it selectively modulates (e.g., inhibits oractivates) PKM2 over at least one other isoform of PK, e.g., it isselective for PKM2 over PKR, PKM1, or PKL; is has an affinity for PKM2which is greater than its affinity for at least one other isoform of PK,e.g., PKR, PKM1, or PKL.

In another embodiment the activator of PKM2 utilized in the methods andcompositions of this invention operates by or has one or more of thefollowing mechanisms or properties:

-   -   a. it is an allosteric activator of PKM2;    -   b. it modulates (e.g., stabilizes or inhibits) the binding of        FBP in a binding pocket of PKM2;    -   c. it modulates (e.g., inhibits or promotes) the release of FBP        from a binding pocket of PKM2;    -   d. it is a modulator (e.g., an agonist or antagonist), e.g., an        analog, of FBP, e.g., an agonist which binds PKM2 with a lower,        about the same, or higher affinity than does FBP;    -   e. it modulates (e.g., inhibits or promotes) the dissolution of        tetrameric PKM2;    -   f. it modulates (e.g., inhibits or promotes) the assembly of        tetrameric PKM2;    -   g. it modulates (e.g., stabilizes or inhibits) the tetrameric        conformation of PKM2;    -   h. it modulates (e.g., inhibits or promotes) the binding of a        phosphotyrosine containing polypeptide to PKM2;    -   i. it modulates (e.g., inhibits or promotes) the ability of a        phosphotyrosine containing polypeptide to induce release of FBP        from PKM2, e.g., by inducing a change in the conformation of        PKM2, e.g., in the position of Lys 433, thereby hindering the        release of FBP;    -   k. it binds to or changes the position of Lys 433 relative to        the FBP binding pocket;    -   l. it selectively modulates (e.g., activates or inhibits) PKM2        over at least one other isoform of PK, e.g., it is selective for        PKM2 over one or more of PKR, PKM1, or PKL;    -   m. it has an affinity for PKM2 which is greater than its        affinity for at least one other isoform of PK, e.g., PKR, PKM1,        or PKL.

A compound described herein may be an activator of PKM2. Exemplarycompounds are shown in Table 1. As shown in Table 1, A refers to anactivator of PKM2 with an EC₅₀<100 nM. B refers to an activator of PKM2with an EC₅₀ between 100 nM and 500 nM. C refers to an activator of PKM2with an EC₅₀ between 500 nM and 1000 nM. D refers to an activator ofPKM2 with an EC₅₀ between 1 μM and 10 μM. E refers to an activator ofPKM2 with an EC₅₀>10 μM ND means not determined.

Ex vivo data is provided as follows: + refers to a compound having anactivity of ≦1 μM; ++ refers to a compound having an activity of >1 μM;ND means not determined

TABLE 1 Structure PKM2_AC50 Ex-Vivo_AC50

A +

D ND

B +

D ND

C ND

B ++

A +

D ND

B ND

C ND

B ND

A +

A +

D ND

D ND

A +

D ND

C ND

E ND

A ++

D ND

C ND

A +

D ND

B ND

A +

A +

D ND

B +

A +

E ND

E ND

A +

C ND

A +

C ND

B ND

B ND

D ND

A ++

A +

B ND

A ND

D ND

D ND

B +

D ND

D ND

C ND

D ND

D ND

B +

B ND

B ND

D ND

D ND

D ND

A +

B ND

E ND

D ND

E ND

A +

D ND

D ND

D ND

D ND

A +

B ND

A +

D ND

C ND

A +

A ND

B +

B ND

B ND

D ND

B ND

E ND

ND ND

D ND

E ND

A +

D ND

C ND

B ND

A +

A +

C ND

D ND

B ND

B +

A +

B ND

B ND

D ND

D ND

D ND

B +

D ND

B +

B +

A ND

D ND

D ND

B ND

B ND

C ND

D ND

C ND

D ND

A ++

B ND

B ND

D +

B ND

C ND

C ND

A ND

A ND

D ND

E ND

D ND

D ND

D ND

B ND

C ND

B ND

A ND

D ND

B ND

B ND

A ND

D ND

D ND

D ND

E ND

D ND

E ND

C ND

D ND

D ND

D ND

C ND

B ND

The compounds described herein can be made using a variety of synthetictechniques. Scheme 1 below depicts a representative synthesis of certaincompounds described herein.

As can be appreciated by the skilled artisan, methods of synthesizingthe compounds of the formulae herein will be evident to those ofordinary skill in the art. Additionally, the various synthetic steps maybe performed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons(1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The compounds of this invention may contain one or more asymmetriccenters and thus occur as racemates and racemic mixtures, singleenantiomers, individual diastereomers and diastereomeric mixtures. Allsuch isomeric forms of these compounds are expressly included in thepresent invention. The compounds of this invention may also containlinkages (e.g., carbon-carbon bonds) or substituents that can restrictbond rotation, e.g. restriction resulting from the presence of a ring ordouble bond. Accordingly, all cis/trans and E/Z isomers are expresslyincluded in the present invention.

The compounds of this invention may also be represented in multipletautomeric forms, in such instances, the invention expressly includesall tautomeric forms of the compounds described herein, even though onlya single tautomeric form may be represented (e.g., alkylation of a ringsystem may result in alkylation at multiple sites, the inventionexpressly includes all such reaction products). All such isomeric formsof such compounds are expressly included in the present invention. Allcrystal forms of the compounds described herein are expressly includedin the present invention including hydrates and other solvates.

The compounds of this invention include the compounds themselves, aswell as their salts and their prodrugs, if applicable. A salt, forexample, can be formed between an anion and a positively chargedsubstituent (e.g., amino) on a compound described herein. Suitableanions include chloride, bromide, iodide, sulfate, nitrate, phosphate,citrate, methanesulfonate, trifluoroacetate, and acetate. Likewise, asalt can also be formed between a cation and a negatively chargedsubstituent (e.g., carboxylate) on a compound described herein. Suitablecations include sodium ion, potassium ion, magnesium ion, calcium ion,and an ammonium cation such as tetramethylammonium ion. Examples ofprodrugs include esters and other pharmaceutically acceptablederivatives, which, upon administration to a subject, are capable ofproviding active compounds.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selected biological properties, e.g.,targeting to a particular tissue. Such modifications are known in theart and include those which increase biological penetration into a givenbiological compartment (e.g., blood, lymphatic system, central nervoussystem), increase oral availability, increase solubility to allowadministration by injection, alter metabolism and alter rate ofexcretion.

In an alternate embodiment, the compounds described herein may be usedas platforms or scaffolds that may be utilized in combinatorialchemistry techniques for preparation of derivatives and/or chemicallibraries of compounds. Such derivatives and libraries of compounds havebiological activity and are useful for identifying and designingcompounds possessing a particular activity. Combinatorial techniquessuitable for utilizing the compounds described herein are known in theart as exemplified by Obrecht, D. and Villalgrodo, J. M.,Solid-Supported Combinatorial and Parallel Synthesis ofSmall-Molecular-Weight Compound Libraries, Pergamon-Elsevier ScienceLimited (1998), and include those such as the “split and pool” or“parallel” synthesis techniques, solid-phase and solution-phasetechniques, and encoding techniques (see, for example, Czarnik, A. W.,Curr. Opin. Chem. Bio., (1997) 1, 60. Thus, one embodiment relates to amethod of using the compounds described herein for generatingderivatives or chemical libraries comprising: 1) providing a bodycomprising a plurality of wells; 2) providing one or more compoundsidentified by methods described herein in each well; 3) providing anadditional one or more chemicals in each well; 4) isolating theresulting one or more products from each well. An alternate embodimentrelates to a method of using the compounds described herein forgenerating derivatives or chemical libraries comprising: 1) providingone or more compounds described herein attached to a solid support; 2)treating the one or more compounds identified by methods describedherein attached to a solid support with one or more additionalchemicals; 3) isolating the resulting one or more products from thesolid support. In the methods described above, “tags” or identifier orlabeling moieties may be attached to and/or detached from the compoundsdescribed herein or their derivatives, to facilitate tracking,identification or isolation of the desired products or theirintermediates. Such moieties are known in the art. The chemicals used inthe aforementioned methods may include, for example, solvents, reagents,catalysts, protecting group and deprotecting group reagents and thelike. Examples of such chemicals are those that appear in the varioussynthetic and protecting group chemistry texts and treatises referencedherein.

Methods of Evaluating Compounds

The compounds described herein can be evaluated for ability to modulatePKM2 (e.g., activate or inhibit PKM2) by methods known in the art.Exemplary methods include contacting the compound with a cell-basedassay which allows assessment of the ability to modulate (e.g., activateor inhibit) PKM2. E.g., the candidate compound can be contacted with acell and measuring the consumption of oxygen or production of lactate. Achange in cellular phosphoenolpyruvate, a change in glycerol-phosphate,a change in ribose or deoxyribose, a change in lipid synthesis, or achange in glucose conversion to lipid or nucleic acids or amino acids orprotein can also be used to evaluate a compound for its ability tomodulate PKM2 (e.g., activate or inhibit PKM2). The evaluation couldalso include measuring a change in pyruvate or a determination of analteration in mitochondrial membrane potential, e.g., as measured byfluorescent potentiometric dyes.

PKM1 and PKM2 for use in the screening method may be produced by anymethod known in the art for expression of recombinant proteins. Forexample, nucleic acids that encode the desired polypeptide may beintroduced into various cell types or cell-free systems for expression.Eukaryotic (e.g., COS, HEK293T, CHO, and NIH cell lines) and prokaryotic(e.g., E. coli) expression systems may be generated in which a PKMsequence is introduced into a plasmid or other vector, which is thenused to transform living cells. Constructs in which the PKM cDNAcontains the entire open reading frame, or biologically active fragmentthereof, are inserted in the correct orientation into an expressionplasmid and may be used for protein expression. Prokaryotic andeukaryotic expression systems allow for the expression and recovery offusion proteins in which the PKM protein is covalently linked to a tagmolecule on either the amino terminal or carboxy terminal side, whichfacilitates identification and/or purification. Examples of tags thatcan be used include hexahistidine, HA, FLAG, and c-myc epitope tags. Anenzymatic or chemical cleavage site can be engineered between the PKMprotein and the tag molecule so that the tag can be removed followingpurification.

The activity of the PKM enzyme measured in the screening assay may bemeasured by, e.g., monitoring the concentration of a substrate (e.g.,ATP or NADH) present in the reaction mixture. Pyruvate, produced by theenzymatic activity of pyruvate kinase, is converted into lactate bylactate dehydrogenase, which requires the consumption of NADH(NADH→NAD+). Thus, the activity of PKM2 can be indirectly measured bymonitoring the consumption of NADH through, e.g., fluorescence assays.Additionally, the activity of the PKM2 enzyme can be directly monitoredby measuring the production of ATP, as ATP is produced whenphosphoenolpyruvate is converted to pyruvate. Methods for monitoring theamount of substrate in a reaction mixture include, e.g., absorbance,fluorescence, Raman scattering, phosphorescence, luminescence,luciferase assays, and radioactivity.

The screening procedure requires the presence of specific components inthe reaction mixture. Components utilized in the assay include, e.g., anucleoside diphosphate (e.g., ADP), phosphoenolpyruvate, NADH, lactatedehydrogenase, FBP, a reducing agent (e.g., dithiothreitol), a detergent(e.g., Brij 35), glycerol, and a solvent (e.g., DMSO). Exemplaryreaction conditions are found in Table 2.

TABLE 2 Component of Amount in Amount in Reaction Condition InhibitionAssay Activation Assay ADP 0.1-5.0 mM 0.1-5.0 mM Phosphoenolpyruvate0.1-5.0 mM 0.1-5.0 mM NADH 10-1000 μM 10-1000 μM Lactate dehydrogenase0.1-10 units 0.1-10 units Fructose-1,6-bisphosphate 1-500 μM 0 DTT0.1-50 mM 0.1-50 mM Brij 35 0.01-1% 0.01-1% Glycerol 0.1-10% 0.1-10%Pyruvate Kinase M2 1-100 pg 1-100 pg (used for screen) DMSO   1-10%  1-10%

Candidate inhibitory compounds are chosen if they demonstratespecificity for PKM2 and inhibition of the PKM2 enzyme greater than 10,15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 99,or 99.9%.

Candidate activator compounds are chosen if they demonstrate specificityand activation of PKM2 enzyme in the absence of FBP to a level greaterthan that of 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80,85, 90, 95, 99, or 100% in the presence of FBP. Furthermore, specificcandidate activators of PKM2 can be evaluated in the presence or absenceof a phosphotyrosine peptide. Phosphotyrosine peptide binding to PKM2leads to a dissociation of FBP from PKM2 and conformational changes ofPKM2 from an active, tetrameric form to an inactive form. Compounds thatbind to PKM2 and lock the enzyme in the active confirmation even in thepresence of a phosphotyrosine peptide will lead to the loss ofallosteric control of PKM2 needed for shunting the biochemicalintermediates from glycolysis into biosynthesis of other intermediates.This, in turn, will lead to inhibition of growth of cancer cells,activated immune cells and fat cells.

Exemplary screening assays also include ex vivo assays, for example, anex vivo assay described herein.

Methods of Treatment

The compounds and compositions described herein can be administered tocells in culture, e.g. in vitro or ex vivo, or to a subject, e.g., invivo, to treat, prevent, and/or diagnose a variety of disorders,including those described herein below.

As used herein, the term “treat” or “treatment” is defined as theapplication or administration of a compound, alone or in combinationwith, a second compound to a subject, e.g., a patient, or application oradministration of the compound to an isolated tissue or cell, e.g., cellline, from a subject, e.g., a patient, who has a disorder (e.g., adisorder as described herein), a symptom of a disorder, or apredisposition toward a disorder, with the purpose to cure, heal,alleviate, relieve, alter, remedy, ameliorate, improve or affect thedisorder, one or more symptoms of the disorder or the predispositiontoward the disorder (e.g., to prevent at least one symptom of thedisorder or to delay onset of at least one symptom of the disorder).

As used herein, an amount of a compound effective to treat a disorder,or a “therapeutically effective amount” refers to an amount of thecompound which is effective, upon single or multiple dose administrationto a subject, in treating a cell, or in curing, alleviating, relievingor improving a subject with a disorder beyond that expected in theabsence of such treatment.

As used herein, an amount of a compound effective to prevent a disorder,or a “a prophylactically effective amount” of the compound refers to anamount effective, upon single- or multiple-dose administration to thesubject, in preventing or delaying the occurrence of the onset orrecurrence of a disorder or a symptom of the disorder.

As used herein, the term “subject” is intended to include human andnon-human animals. Exemplary human subjects include a human patienthaving a disorder, e.g., a disorder described herein or a normalsubject. The term “non-human animals” of the invention includes allvertebrates, e.g., non-mammals (such as chickens, amphibians, reptiles)and mammals, such as non-human primates, domesticated and/oragriculturally useful animals, e.g., sheep, dog, cat, cow, pig, etc.

Neoplastic Disorders

A compound or composition described herein can be used to treat aneoplastic disorder. A “neoplastic disorder” is a disease or disordercharacterized by cells that have the capacity for autonomous growth orreplication, e.g., an abnormal state or condition characterized byproliferative cell growth. Exemplary neoplastic disorders include:carcinoma, sarcoma, metastatic disorders (e.g., tumors arising fromprostate, colon, lung, breast and liver origin), hematopoieticneoplastic disorders, e.g., leukemias, metastatic tumors. Prevalentcancers include: breast, prostate, colon, lung, liver, and pancreaticcancers. Treatment with the compound may be in an amount effective toameliorate at least one symptom of the neoplastic disorder, e.g.,reduced cell proliferation, reduced tumor mass, etc.

The disclosed methods are useful in the prevention and treatment ofcancer, including for example, solid tumors, soft tissue tumors, andmetastases thereof. The disclosed methods are also useful in treatingnon-solid cancers. Exemplary solid tumors include malignancies (e.g.,sarcomas, adenocarcinomas, and carcinomas) of the various organ systems,such as those of lung, breast, lymphoid, gastrointestinal (e.g., colon),and genitourinary (e.g., renal, urothelial, or testicular tumors)tracts, pharynx, prostate, and ovary. Exemplary adenocarcinomas includecolorectal cancers, renal-cell carcinoma, liver cancer, non-small cellcarcinoma of the lung, and cancer of the small intestine.

Exemplary cancers described by the national cancer institute include:Acute Lymphoblastic Leukemia, Adult; Acute Lymphoblastic Leukemia,Childhood; Acute Myeloid Leukemia, Adult; Adrenocortical Carcinoma;Adrenocortical Carcinoma, Childhood; AIDS-Related Lymphoma; AIDS-RelatedMalignancies; Anal Cancer; Astrocytoma, Childhood Cerebellar;Astrocytoma, Childhood Cerebral; Bile Duct Cancer, Extrahepatic; BladderCancer; Bladder Cancer, Childhood; Bone Cancer, Osteosarcoma/MalignantFibrous Histiocytoma; Brain Stem Glioma, Childhood; Brain Tumor, Adult;Brain Tumor, Brain Stem Glioma, Childhood; Brain Tumor, CerebellarAstrocytoma, Childhood; Brain Tumor, Cerebral Astrocytoma/MalignantGlioma, Childhood; Brain Tumor, Ependymoma, Childhood; Brain Tumor,Medulloblastoma, Childhood; Brain Tumor, Supratentorial PrimitiveNeuroectodermal Tumors, Childhood; Brain Tumor, Visual Pathway andHypothalamic Glioma, Childhood; Brain Tumor, Childhood (Other); BreastCancer; Breast Cancer and Pregnancy; Breast Cancer, Childhood; BreastCancer, Male; Bronchial Adenomas/Carcinoids, Childhood; Carcinoid Tumor,Childhood; Carcinoid Tumor, Gastrointestinal; Carcinoma, Adrenocortical;Carcinoma, Islet Cell; Carcinoma of Unknown Primary; Central NervousSystem Lymphoma, Primary; Cerebellar Astrocytoma, Childhood; CerebralAstrocytoma/Malignant Glioma, Childhood; Cervical Cancer; ChildhoodCancers; Chronic Lymphocytic Leukemia; Chronic Myelogenous Leukemia;Chronic Myeloproliferative Disorders; Clear Cell Sarcoma of TendonSheaths; Colon Cancer; Colorectal Cancer, Childhood; Cutaneous T-CellLymphoma; Endometrial Cancer; Ependymoma, Childhood; Epithelial Cancer,Ovarian; Esophageal Cancer; Esophageal Cancer, Childhood; Ewing's Familyof Tumors; Extracranial Germ Cell Tumor, Childhood; Extragonadal GermCell Tumor; Extrahepatic Bile Duct Cancer; Eye Cancer, IntraocularMelanoma; Eye Cancer, Retinoblastoma; Gallbladder Cancer; Gastric(Stomach) Cancer; Gastric (Stomach) Cancer, Childhood; GastrointestinalCarcinoid Tumor; Germ Cell Tumor, Extracranial, Childhood; Germ CellTumor, Extragonadal; Germ Cell Tumor, Ovarian; Gestational TrophoblasticTumor; Glioma, Childhood Brain Stem; Glioma, Childhood Visual Pathwayand Hypothalamic; Hairy Cell Leukemia; Head and Neck Cancer;Hepatocellular (Liver) Cancer, Adult (Primary); Hepatocellular (Liver)Cancer, Childhood (Primary); Hodgkin's Lymphoma, Adult; Hodgkin'sLymphoma, Childhood; Hodgkin's Lymphoma During Pregnancy; HypopharyngealCancer; Hypothalamic and Visual Pathway Glioma, Childhood; IntraocularMelanoma; Islet Cell Carcinoma (Endocrine Pancreas); Kaposi's Sarcoma;Kidney Cancer; Laryngeal Cancer; Laryngeal Cancer, Childhood; Leukemia,Acute Lymphoblastic, Adult; Leukemia, Acute Lymphoblastic, Childhood;Leukemia, Acute Myeloid, Adult; Leukemia, Acute Myeloid, Childhood;Leukemia, Chronic Lymphocytic; Leukemia, Chronic Myelogenous; Leukemia,Hairy Cell; Lip and Oral Cavity Cancer; Liver Cancer, Adult (Primary);Liver Cancer, Childhood (Primary); Lung Cancer, Non-Small Cell; LungCancer, Small Cell; Lymphoblastic Leukemia, Adult Acute; LymphoblasticLeukemia, Childhood Acute; Lymphocytic Leukemia, Chronic; Lymphoma,AIDS-Related; Lymphoma, Central Nervous System (Primary); Lymphoma,Cutaneous T-Cell; Lymphoma, Hodgkin's, Adult; Lymphoma, Hodgkin's,Childhood; Lymphoma, Hodgkin's During Pregnancy; Lymphoma,Non-Hodgkin's, Adult; Lymphoma, Non-Hodgkin's, Childhood; Lymphoma,Non-Hodgkin's During Pregnancy; Lymphoma, Primary Central NervousSystem; Macroglobulinemia, Waldenstrom's; Male Breast Cancer; MalignantMesothelioma, Adult; Malignant Mesothelioma, Childhood; MalignantThymoma; Medulloblastoma, Childhood; Melanoma; Melanoma, Intraocular;Merkel Cell Carcinoma; Mesothelioma, Malignant; Metastatic Squamous NeckCancer with Occult Primary; Multiple Endocrine Neoplasia Syndrome,Childhood; Multiple Myeloma/Plasma Cell Neoplasm; Mycosis Fungoides;Myelodysplastic Syndromes; Myelogenous Leukemia, Chronic; MyeloidLeukemia, Childhood Acute; Myeloma, Multiple; MyeloproliferativeDisorders, Chronic; Nasal Cavity and Paranasal Sinus Cancer;Nasopharyngeal Cancer; Nasopharyngeal Cancer, Childhood; Neuroblastoma;Non-Hodgkin's Lymphoma, Adult; Non-Hodgkin's Lymphoma, Childhood;Non-Hodgkin's Lymphoma During Pregnancy; Non-Small Cell Lung Cancer;Oral Cancer, Childhood; Oral Cavity and Lip Cancer; OropharyngealCancer; Osteosarcoma/Malignant Fibrous Histiocytoma of Bone; OvarianCancer, Childhood; Ovarian Epithelial Cancer; Ovarian Germ Cell Tumor;Ovarian Low Malignant Potential Tumor; Pancreatic Cancer; PancreaticCancer, Childhood; Pancreatic Cancer, Islet Cell; Paranasal Sinus andNasal Cavity Cancer; Parathyroid Cancer; Penile Cancer;Pheochromocytoma; Pineal and Supratentorial Primitive NeuroectodermalTumors, Childhood; Pituitary Tumor; Plasma Cell Neoplasm/MultipleMyeloma; Pleuropulmonary Blastoma; Pregnancy and Breast Cancer;Pregnancy and Hodgkin's Lymphoma; Pregnancy and Non-Hodgkin's Lymphoma;Primary Central Nervous System Lymphoma; Primary Liver Cancer, Adult;Primary Liver Cancer, Childhood; Prostate Cancer; Rectal Cancer; RenalCell (Kidney) Cancer; Renal Cell Cancer, Childhood; Renal Pelvis andUreter, Transitional Cell Cancer; Retinoblastoma; Rhabdomyosarcoma,Childhood; Salivary Gland Cancer; Salivary Gland Cancer, Childhood;Sarcoma, Ewing's Family of Tumors; Sarcoma, Kaposi's; Sarcoma(Osteosarcoma)/Malignant Fibrous Histiocytoma of Bone; Sarcoma,Rhabdomyosarcoma, Childhood; Sarcoma, Soft Tissue, Adult; Sarcoma, SoftTissue, Childhood; Sezary Syndrome; Skin Cancer; Skin Cancer, Childhood;Skin Cancer (Melanoma); Skin Carcinoma, Merkel Cell; Small Cell LungCancer; Small Intestine Cancer; Soft Tissue Sarcoma, Adult; Soft TissueSarcoma, Childhood; Squamous Neck Cancer with Occult Primary,Metastatic; Stomach (Gastric) Cancer; Stomach (Gastric) Cancer,Childhood; Supratentorial Primitive Neuroectodermal Tumors, Childhood;T-Cell Lymphoma, Cutaneous; Testicular Cancer; Thymoma, Childhood;Thymoma, Malignant; Thyroid Cancer; Thyroid Cancer, Childhood;Transitional Cell Cancer of the Renal Pelvis and Ureter; TrophoblasticTumor, Gestational; Unknown Primary Site, Cancer of, Childhood; UnusualCancers of Childhood; Ureter and Renal Pelvis, Transitional Cell Cancer;Urethral Cancer; Uterine Sarcoma; Vaginal Cancer; Visual Pathway andHypothalamic Glioma, Childhood; Vulvar Cancer; Waldenstrom's Macroglobulinemia; and Wilms' Tumor. Metastases of the aforementioned cancerscan also be treated or prevented in accordance with the methodsdescribed herein.

Cancer Combination Therapies

In some embodiments, a compound described herein is administeredtogether with an additional cancer treatment. Exemplary cancertreatments include, for example: chemotherapy, targeted therapies suchas antibody therapies, immunotherapy, and hormonal therapy. Examples ofeach of these treatments are provided below.

Chemotherapy

In some embodiments, a compound described herein is administered with achemotherapy. Chemotherapy is the treatment of cancer with drugs thatcan destroy cancer cells. “Chemotherapy” usually refers to cytotoxicdrugs which affect rapidly dividing cells in general, in contrast withtargeted therapy. Chemotherapy drugs interfere with cell division invarious possible ways, e.g., with the duplication of DNA or theseparation of newly formed chromosomes. Most forms of chemotherapytarget all rapidly dividing cells and are not specific for cancer cells,although some degree of specificity may come from the inability of manycancer cells to repair DNA damage, while normal cells generally can.

Examples of chemotherapeutic agents used in cancer therapy include, forexample, antimetabolites (e.g., folic acid, purine, and pyrimidinederivatives) and alkylating agents (e.g., nitrogen mustards,nitrosoureas, platinum, alkyl sulfonates, hydrazines, triazenes,aziridines, spindle poison, cytotoxic agents, toposimerase inhibitorsand others). Exemplary agents include Aclarubicin, Actinomycin,Alitretinon, Altretamine, Aminopterin, Aminolevulinic acid, Amrubicin,Amsacrine, Anagrelide, Arsenic trioxide, Asparaginase, Atrasentan,Belotecan, Bexarotene, endamustine, Bleomycin, Bortezomib, Busulfan,Camptothecin, Capecitabine, Carboplatin, Carboquone, Carmofur,Carmustine, Celecoxib, Chlorambucil, Chlormethine, Cisplatin,Cladribine, Clofarabine, Crisantaspase, Cyclophosphamide, Cytarabine,Dacarbazine, Dactinomycin, Daunorubicin, Decitabine, Demecolcine,Docetaxel, Doxorubicin, Efaproxiral, Elesclomol, Elsamitrucin,Enocitabine, Epirubicin, Estramustine, Etoglucid, Etoposide,Floxuridine, Fludarabine, Fluorouracil (5FU), Fotemustine, Gemcitabine,Gliadel implants, Hydroxycarbamide, Hydroxyurea, Idarubicin, Ifosfamide,Irinotecan, Irofulven, Ixabepilone, Larotaxel, Leucovorin, Liposomaldoxorubicin, Liposomal daunorubicin, Lonidamine, Lomustine, Lucanthone,Mannosulfan, Masoprocol, Melphalan, Mercaptopurine, Mesna, Methotrexate,Methyl aminolevulinate, Mitobronitol, Mitoguazone, Mitotane, Mitomycin,Mitoxantrone, Nedaplatin, Nimustine, Oblimersen, Omacetaxine, Ortataxel,Oxaliplatin, Paclitaxel, Pegaspargase, Pemetrexed, Pentostatin,Pirarubicin, Pixantrone, Plicamycin, Porfimer sodium, Prednimustine,Procarbazine, Raltitrexed, Ranimustine, Rubitecan, Sapacitabine,Semustine, Sitimagene ceradenovec, Strataplatin, Streptozocin,Talaporfin, Tegafur-uracil, Temoporfin, Temozolomide, Teniposide,Tesetaxel, Testolactone, Tetranitrate, Thiotepa, Tiazofurine,Tioguanine, Tipifarnib, Topotecan, Trabectedin, Triaziquone,Triethylenemelamine, Triplatin, Tretinoin, Treosulfan, Trofosfamide,Uramustine, Valrubicin, Verteporfin, Vinblastine, Vincristine,Vindesine, Vinflunine, Vinorelbine, Vorinostat, Zorubicin, and othercytostatic or cytotoxic agents described herein.

Because some drugs work better together than alone, two or more drugsare often given at the same time. Often, two or more chemotherapy agentsare used as combination chemotherapy. In some embodiments, thechemotherapy agents (including combination chemotherapy) can be used incombination with a compound described herein.

Targeted Therapy

In some embodiments, a compound described herein is administered with atargeted therapy. Targeted therapy constitutes the use of agentsspecific for the deregulated proteins of cancer cells. Small moleculetargeted therapy drugs are generally inhibitors of enzymatic domains onmutated, overexpressed, or otherwise critical proteins within the cancercell. Prominent examples are the tyrosine kinase inhibitors such asAxitinib, Bosutinib, Cediranib, desatinib, erolotinib, imatinib,gefitinib, lapatinib, Lestaurtinib, Nilotinib, Semaxanib, Sorafenib,Sunitinib, and Vandetanib, and also cyclin-depdendent kinase inhibitorssuch as Alvocidib and Seliciclib. Monoclonal antibody therapy is anotherstrategy in which the therapeutic agent is an antibody whichspecifically binds to a protein on the surface of the cancer cells.Examples include the anti-HER2/neu antibody trastuzumab (HERCEPTIN®)typically used in breast cancer, and the anti-CD20 antibody rituximaband Tositumomab typically used in a variety of B-cell malignancies.Other exemplary anbitodies include Ctuximab, Panitumumab, Trastuzumab,Alemtuzumab, Bevacizumab, Edrecolomab, and Gemtuzumab. Exemplary fusionproteins include Aflibercept and Denileukin diftitox. In someembodiments, the targeted therapy can be used in combination with acompound described herein.

Targeted therapy can also involve small peptides as “homing devices”which can bind to cell surface receptors or affected extracellularmatrix surrounding the tumor. Radionuclides which are attached to thesepeptides (e.g., RGDs) eventually kill the cancer cell if the nuclidedecays in the vicinity of the cell. An example of such therapy includesBEXXAR®.

Immunotherapy

In some embodiments, a compound described herein is administered with animmunotherapy. Cancer immunotherapy refers to a diverse set oftherapeutic strategies designed to induce the patient's own immunesystem to fight the tumor. Contemporary methods for generating an immuneresponse against tumors include intravesicular BCG immunotherapy forsuperficial bladder cancer, and use of interferons and other cytokinesto induce an immune response in renal cell carcinoma and melanomapatients.

Allogeneic hematopoietic stem cell transplantation can be considered aform of immunotherapy, since the donor's immune cells will often attackthe tumor in a graft-versus-tumor effect. In some embodiments, theimmunotherapy agents can be used in combination with a compounddescribed herein.

Hormonal Therapy

In some embodiments, a compound described herein is administered with ahormonal therapy. The growth of some cancers can be inhibited byproviding or blocking certain hormones. Common examples ofhormone-sensitive tumors include certain types of breast and prostatecancers. Removing or blocking estrogen or testosterone is often animportant additional treatment. In certain cancers, administration ofhormone agonists, such as progestogens may be therapeuticallybeneficial. In some embodiments, the hormonal therapy agents can be usedin combination with a compound described herein.

Obesity and Fat Disorders

A compound or composition described herein can be used to treat orprevent obesity, e.g., in a human subject, e.g. a child or adultsubject. “Obesity” refers to a condition in which a subject has a bodymass index of greater than or equal to 30. Many compounds describedherein can be used to treat or prevent an over-weight condition.“Over-weight” refers to a condition in which a subject has a body massindex of greater or equal to 25.0. The body mass index (BMI) and otherdefinitions are according to the “NIH Clinical Guidelines on theIdentification and Evaluation, and Treatment of Overweight and Obesityin Adults” (1998). Treatment with the compound may be in an amounteffective to alter the weight of the subject, e.g., by at least 2, 5, 7,10, 12, 15, 20, 25, 30, 25, 40, 45, 50, or 55%. Treatment with acompound may be in an amount effective to reduce the body mass index ofthe subject, e.g., to less than 30, 28, 27, 25, 22, 20, or 18. Thecompounds can be used to treat or prevent aberrant or inappropriateweight gain, metabolic rate, or fat deposition, e.g., anorexia, bulimia,obesity, diabetes, or hyperlipidemia (e.g., elevated triglyceridesand/or elevated cholesterol), as well as disorders of fat or lipidmetabolism.

A compound or composition described herein can be administered to treatobesity associated with Prader-Willi Syndrome (PWS). PWS is a geneticdisorder associated with obesity (e.g., morbid obesity).

A compound or composition described herein can be used to reduce bodyfat, prevent increased body fat, reduce cholesterol (e.g., totalcholesterol and/or ratios of total cholesterol to HDL cholesterol),and/or reduce appetite in individuals having PWS associated obesity,and/or reduce comorbidities such as diabetes, cardiovascular disease,and stroke.

Compositions and Routes of Administration

The compositions delineated herein include the compounds delineatedherein (e.g., a compound described herein), as well as additionaltherapeutic agents if present, in amounts effective for achieving amodulation of disease or disease symptoms, including those describedherein.

The term “pharmaceutically acceptable carrier or adjuvant” refers to acarrier or adjuvant that may be administered to a patient, together witha compound of this invention, and which does not destroy thepharmacological activity thereof and is nontoxic when administered indoses sufficient to deliver a therapeutic amount of the compound.

Pharmaceutically acceptable carriers, adjuvants and vehicles that may beused in the pharmaceutical compositions of this invention include, butare not limited to, ion exchangers, alumina, aluminum stearate,lecithin, self-emulsifying drug delivery systems (SEDDS) such asd-α-tocopherol polyethyleneglycol 1000 succinate, surfactants used inpharmaceutical dosage forms such as Tweens or other similar polymericdelivery matrices, serum proteins, such as human serum albumin, buffersubstances such as phosphates, glycine, sorbic acid, potassium sorbate,partial glyceride mixtures of saturated vegetable fatty acids, water,salts or electrolytes, such as protamine sulfate, disodium hydrogenphosphate, potassium hydrogen phosphate, sodium chloride, zinc salts,colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone,cellulose-based substances, polyethylene glycol, sodiumcarboxymethylcellulose, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat. Cyclodextrins such as α-, β-, and γ-cyclodextrin, orchemically modified derivatives such as hydroxyalkylcyclodextrins,including 2- and 3-hydroxypropyl-β-cyclodextrins, or other solubilizedderivatives may also be advantageously used to enhance delivery ofcompounds of the formulae described herein.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intraarticular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, or carboxymethyl cellulose or similar dispersing agentswhich are commonly used in the formulation of pharmaceuticallyacceptable dosage forms such as emulsions and or suspensions. Othercommonly used surfactants such as Tweens or Spans and/or other similaremulsifying agents or bioavailability enhancers which are commonly usedin the manufacture of pharmaceutically acceptable solid, liquid, orother dosage forms may also be used for the purposes of formulation.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, emulsions and aqueous suspensions,dispersions and solutions. In the case of tablets for oral use, carrierswhich are commonly used include lactose and corn starch. Lubricatingagents, such as magnesium stearate, are also typically added. For oraladministration in a capsule form, useful diluents include lactose anddried corn starch. When aqueous suspensions and/or emulsions areadministered orally, the active ingredient may be suspended or dissolvedin an oily phase is combined with emulsifying and/or suspending agents.If desired, certain sweetening and/or flavoring and/or coloring agentsmay be added.

The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature and therefore willmelt in the rectum to release the active components. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

Topical administration of the pharmaceutical compositions of thisinvention is useful when the desired treatment involves areas or organsreadily accessible by topical application. For application topically tothe skin, the pharmaceutical composition should be formulated with asuitable ointment containing the active components suspended ordissolved in a carrier. Carriers for topical administration of thecompounds of this invention include, but are not limited to, mineraloil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier with suitable emulsifying agents. Suitablecarriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol,2-octyldodecanol, benzyl alcohol and water. The pharmaceuticalcompositions of this invention may also be topically applied to thelower intestinal tract by rectal suppository formulation or in asuitable enema formulation. Topically-transdermal patches are alsoincluded in this invention.

The pharmaceutical compositions of this invention may be administered bynasal aerosol or inhalation. Such compositions are prepared according totechniques well-known in the art of pharmaceutical formulation and maybe prepared as solutions in saline, employing benzyl alcohol or othersuitable preservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other solubilizing or dispersing agents known inthe art.

When the compositions of this invention comprise a combination of acompound of the formulae described herein and one or more additionaltherapeutic or prophylactic agents, both the compound and the additionalagent should be present at dosage levels of between about 1 to 100%, andmore preferably between about 5 to 95% of the dosage normallyadministered in a monotherapy regimen. The additional agents may beadministered separately, as part of a multiple dose regimen, from thecompounds of this invention. Alternatively, those agents may be part ofa single dosage form, mixed together with the compounds of thisinvention in a single composition.

The compounds described herein can, for example, be administered byinjection, intravenously, intraarterially, subdermally,intraperitoneally, intramuscularly, or subcutaneously; or orally,buccally, nasally, transmucosally, topically, in an ophthalmicpreparation, or by inhalation, with a dosage ranging from about 0.5 toabout 100 mg/kg of body weight, alternatively dosages between 1 mg and1000 mg/dose, every 4 to 120 hours, or according to the requirements ofthe particular drug. The methods herein contemplate administration of aneffective amount of compound or compound composition to achieve thedesired or stated effect. Typically, the pharmaceutical compositions ofthis invention will be administered from about 1 to about 6 times perday or alternatively, as a continuous infusion. Such administration canbe used as a chronic or acute therapy. The amount of active ingredientthat may be combined with the carrier materials to produce a singledosage form will vary depending upon the host treated and the particularmode of administration. A typical preparation will contain from about 5%to about 95% active compound (w/w). Alternatively, such preparationscontain from about 20% to about 80% active compound.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Patients may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

Patient Selection and Monitoring

The compounds described herein can modulate PKM2. Accordingly, a patientand/or subject can be selected for treatment using a compound describedherein by first evaluating the patient and/or subject to determinewhether the subject is in need of modulation of PKM2, and if the subjectis determined to be in need of modulation of PKM2, then optionallyadministering to the subject a compound described herein.

A subject can be evaluated as being in need of modulation of PKM2 usingmethods known in the art, e.g., by measuring the presence and/oractivity of PKM2 in the patient. In some embodiments, the activityand/or level of PKM2 is evaluated in the cancer.

A patient receiving a compound described herein can be monitored, forexample, for improvement in the condition and/or adverse effects.Improvement of a patient's condition can be evaluated, for example, bymonitoring the growth, absence of growth, or regression of the cancer(e.g., a tumor). In some embodiments, the patient is evaluated using aradiological assay or evaluation of hemolytic parameters.

EXAMPLES Example 1 PKM2 Assay

Procedure:

-   -   PKM2 stock enzyme solution was diluted in Reaction Buffer    -   2 μL of compound was added into each well first, and then 180 μL        of the Reaction Mix was added.    -   Reaction mixture with compound (without ADP) were incubated for        30 minutes at 4° C.    -   Plates were re-equilibrated to room temperature prior to adding        20 μL ADP to initiate the reaction.    -   Reaction progress was measured as changes in absorbance at 340        nm wavelength at room temperature (25° C.)        Reaction Mix: PKM2 (50 ng/well), ADP (0.7 mM), PEP (0.15 mM),        NADH (180 μM), LDH (2 units) in Reaction Buffer        Reaction Buffer: 100 mM KCl, 50 mM Tris pH 7.5, 5 mM MgCl2, 1 mM        DTT, 0.03% BSA.

Example 2 PKM2 Ex-Vivo Assay

Described herein is a method to measure the activity of PKM2 activatorsin living cells and tissue. One of ordinary skill in the art wouldrecognize and understand that this method can be adapted to highthroughput format, and can accommodate a variety of cell lines andgrowth conditions.

In the assay, cells are treated with a compound described herein (i.e.,a PKM2 activator). This compound is capable of entering the cell andbinding to PKM2, inducing an activated conformation. The excess unboundcompound is washed away with PBS, and the cells are lysed bysnap-freezing on dry ice, followed by addition of a detergent-containinglysis buffer. The lysate, in which activated PKM2 remains intact, isremoved and added to a chemical cocktail including the chemicalsnecessary to measure pyruvate kinase activity, in an assay that iscoupled to the LDHa enzyme. The amount of pyruvate kinase activity thatis measured is normalized to the total protein content in the lysate,and related to the concentration of PKM2 activator that was added to thecell. This allows an AC₅₀ (concentration at which PKM2 is activated 50%)value to be derived. The total fold-increase in activity overmock-treated cells can also be calculated, and the “maximum level ofactivation” can be used to distinguish between compounds that fullyactivate PKM2 and compounds that can only partially activate PKM2.

In the case of measuring PKM2 activity from tissue (for example, in acell tumor), animals harboring the tissue/tumor of interest are dosedwith a compound. After a specified period of time in which exposure hasbeen achieved in the target tissue/tumor of interest, the tissue/tumoris harvested from the animal, snap-frozen, and then lysed andhomogenized. The amount of pyruvate kinase activity in this lysate canthen be quantitated as described above.

Materials:

-   Lysis buffer* * This lysis buffer (without PMSF) is available from    Cell Signaling Technology as a 10× stock (#9803)-   20 mM Tris-HCl (pH 7.5)-   150 mM NaCl-   1 mM Na₂EDTA-   1 mM EGTA-   1% Triton-   2.5 mM sodium pyrophosphate-   1 mM beta-glycerophosphate-   1 mM Na₃VO₄-   1 μg/ml leupeptin-   1 mM PMSF** 1 mM PMSF is added fresh from a 100 mM stock solution    made up in isopropanol. The stock solution can be stored at 4    degrees indefinitely.    Pyruvate kinase assay master mix (Same for PKM2 Activator Assay):

TABLE 3 KCl 100 mM Tris (pH 7.5) 50 mM MgCl₂ 5.0 mM PEP 0.10 mM NADH0.18 mM DTT 1.00 mM BSA 0.3 mg/mL LDH 0.5 units H₂O to 180 uL

ADP Solution:

ADP 7.0 mM H₂O to 20 uLProcedure:

On the first day (day 1) cells are normally cultured in RPMI-1640 (Lonza#12-115° F.) (with 25 mM Hepes, L-glutamine)/10% FBS. The cells aresubsequently trypsinized and plated in RPMI-1640 (Lonza, #12-918F) (nophenol red, supplemented with L-glutamine @300 mg/L (Sigma, #G8540))/10%FBS at the following densities in 96 well plates:

A549: 40 k/well

100 uL final volume of media per well.

On the second day (Day 2), the cells should be 70-90% confluent. Thecells are then treated with a compound described herein dissolved inmedia at final assay concentrations in a 96-well assay block (500 uL)(Costar, #3956). The final DMSO concentration is 0.1% (0.5 μL into 500uL). Compound dilutions in DMSO are prepared so that the final DMSOconcentration is constant at all compound concentrations. The media forthe assay is RPMI-1640 (no phenol red, with L-glutamine @300 mg/L).

The media is then aspirated carefully from the cells using amulti-channel aspirator. 100 μL of media w/compounds is added onto cellswith a multichannel pipette. Each compound concentration is then assayedin triplicate (a duplicate assay is also sufficient).

The cells are treated for 1-4 hrs (this time is determined empiricallycompared to DMSO reference treatment). During the cell treatment, PBS(containing calcium and magnesium) and lysis buffer is cooled on ice.

The cells are lysed and the pyruvate kinase activity is assayed. Theremaining media is aspirated and the cells are washed 2× with 100 uLice-cold PBS. The PBS is removed, and the cell plate frozen on dry icefor 5 minutes. The cells are lysed in 50 L cold lysis buffer. Cells aresubsequently kept on ice for 5 minutes, and then agitated on a plateshaker for 5 minutes (repeat 3×). Remove 10 L for protein quantitation(or use OD₂₈₀ on entire plate).

In a fresh plate, 170 uL of pyruvate kinase assay master mix was addedto each well (see end for recipe). 10 uL of cell lysate was thentransferred into each well. The assay was initiated upon addition of 20uL of ADP solution. The rates were then calculated against the initialrates to determine pyruvate kinase specific activity.

The concentration and type of detergent in the lysis buffer can bevaried to accommodate the specific physicochemical properties of thespecific PKM2 activator. For instance, the interaction between some PKM2activators and PKM2 can be disrupted by higher detergent concentrations,but preserved when cells are lysed with lower detergent concentrations.

Example 3 Compounds and Synthesis

General Procedure for Compound 2: Chlorosulfonic acid (24 mL) was addedslowly over a period of 15 min to imidazole (4.0 g, 58.82 mmol) at 0° C.The resulting mixture was heated at 120-130° C. for 3 h. Aftercompletion of SM, the reaction mixture was quenched with ice cold water(25 mL) and extracted with DCM (3×40 mL). The combined organic layerswere dried over Na₂SO₄ and concentrated under reduced pressure. Thedesired Compound-2 was obtained as a solid (3.0 g, 42% yield).

¹H NMR (500 MHz, DMSO-d₆) δ: 14.29 (bs, 1H), 8.90 (s, 1H), 7.64 (s, 1H);Mass (M+1): 166.9.

General Procedure for Compound 4: To a solution of compound-3 (500 mg,4.13 mmol) in DCM (10 mL) under a nitrogen atmosphere, pyridine (0.65ml, 8.26 mmol) was added and stirred at room temperature for 15 min. Thereaction mixture was then cooled to 0° C. followed by the dropwiseaddition of Compound-2 (820 mg, 4.96 mmol) in DCM (4 mL). The resultingreaction mixture was stirred for 2 h at room temperature. Aftercompletion of the reaction, 0.5N HCl solution was added and extractedwith DCM (2×10 mL). The combined organic layers were washed with brine(1×20 mL), dried over Na₂SO₄ and concentrated under reduced pressure toprovide the desired compound 4 as a solid (500 mg, 50% yield).

General Procedure for Compound 6: In a two neck RB flask, compound-4 (50mg, 0.2 mmoL), compound-5 (51 mg, 0.24 mmoL) and K₂CO₃ (54 mg, 0.4 mmoL)were charged in DMF (6 mL) under N₂ atmosphere. The resulting reactionmixture was stirred at 90° C. for 3 h. The progress of the reaction wasmonitored by TLC. After completion of SM, the reaction mixture wasquenched with water (15 mL) and extracted with ethyl acetate (2×20 mL).The combined organic layers were dried over Na₂SO₄ and concentratedunder reduced pressure. The crude product was purified by columnchromatography (Silica gel 60-120, 3:7, ethyl acetate/hexane) to obtainthe desired compound-6 as a solid (40 mg, 47% yield).

¹H NMR (500 MHz, DMSO-D6) δ: 10.29 (s, 1H), 9.03 (s, 1H), 8.84 (s, 1H),8.37 (s, 1H), 8.32 (s, 1H), 7.82 (s, 2H), 6.63 (s, 1H), 2.18 (s, 6H);HPLC Purity: 93.13%; Mass (M+1): 431.1.

1-(3-chloro-5-(trifluromethyl)pyridine-2-yl)-N-(3,5-dimethylphenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.29 (s, 1H), 9.03 (s, 1H), 8.84 (s, 1H),8.37 (s, 1H), 8.32 (s, 1H), 7.82 (s, 2H), 6.63 (s, 1H), 2.18 (s, 6H);HPLC Purity: 93.13%; Mass (M+1): 431.1.

1-(3-chloro-5-(trifluromethyl)pyridine-2-yl)-N-(4-methoxyphenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.04 (bs, 1H), 9.01 (s, 1H), 8.83 (s, 1H),8.38 (s, 1H), 8.18 (s, 1H), 7.08 (d, 2H), 6.82 (d, 2H), 3.67 (s, 3H);HPLC Purity: 91.04%; Mass (M+1): 433.

N-(4-methoxyphenyl)-1-5-(trifluromethyl)pyridine-2-yl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.11 (s, 1H), 8.97 (s, 1H), 8.81 (s, 1H),8.52 (m, 2H), 8.19 (d, 1H), 7.48 (d, 2H), 6.82 (d, 2H), 3.65 (s, 3H);HPLC Purity: 96.12%; Mass (M+1): 399.

N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-5-(trifluromethyl)pyridine-2-yl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.53 (s, 1H), 8.96 (s, 1H), 8.79 (s, 1H),8.58 (s, 1H), 8.56 (s, 1H), 8.18 (d, 1H), 6.66 (s, 2H), 6.63 (d, 1H)4.17 (t, 4H); HPLC Purity: 98.48%; Mass (M+1): 427.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.13 (s, 1H), 9.02 (s, 1H), 8.84 (s, 1H),8.39 (s, 1H), 8.22 (s, 1H), 6.78-6.58 (m, 3H), 4.18 (s, 4H); HPLCPurity: 96.98%; Mass (M+1): 461.

N-(4-fluorophenyl)-1-(5-(trifluromethyl)pyridine-2-yl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.42 (s, 1H), 8.97 (s, 1H), 8.80 (s, 1H),8.62 (s, 1H), 8.46 (d, 1H), 8.19 (d, 1H), 7.23 (d, 2H), 7.08 (d, 2H);HPLC Purity: 99.35%; Mass (M+1): 387.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(4-fluorophenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.42 (s, 1H), 9.02 (s, 1H), 8.84 (s, 1H),8.39 (s, 1H), 8.24 (s, 1H), 7.22 (d, 2H), 7.08 (d, 2H); HPLC Purity:92.78%; Mass (M+1): 421.

N-(3,5-dimethylphenyl)-1-(5-(trifluromethyl)pyridine-2-yl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.28 (s, 1H), 8.96 (s, 1H), 8.78 (s, 1H),8.63 (s, 1H), 8.33 (d, 1H), 8.19 (d, 1H), 7.82 (s, 2H), 7.62 (s, 1H),2.18 (s, 6H); HPLC Purity: 99.23%; Mass (M+1): 397.1.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(3,5-dimethoxyphenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.41 (s, 1H), 9.02 (s, 1H), 8.87 (s, 1H),8.38 (s, 2H), 6.39 (s, 2H), 6.17 (s, 1H), 3.67 (s, 6H); HPLC Purity:97.82%; Mass (M+1): 463.1.

N-(4-chloro-3-methylphenyl)-1-(3-chloro-(5-(trifluromethyl)pyridine-2-yl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.53 (s, 1H), 9.03 (s, 1H), 8.84 (s, 1H),8.37 (d, 2H), 7.28 (d, 1H), 7.16 (s, 1H), 7.03 (d, 1H), 2.22 (s, 3H);HPLC Purity: 98.76%; Mass (M+1): 451.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(3,4-dimethylphenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.21 (s, 1H), 9.02 (s, 1H), 8.85 (s, 1H),8.38 (s, 2H), 8.24 (s, 1H), 6.98-6.87 (m, 3H), 2.13 (s, 6H); HPLCPurity: 99.06%; Mass (M+1): 431.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(3-chlorophenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.78 (s, 1H), 9.03 (s, 1H), 8.84 (s, 1H),8.38 (d, 2H), 7.31-7.24 (m, 2H), 7.16 (d, 1H), 7.08 (d, 1H); HPLCPurity: 95.26%; Mass (M+1): 436.9.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(3-(trifluromethyl)phenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.96 (s, 1H), 9.02 (s, 1H), 8.87 (s, 1H),8.38 (d, 2H), 7.52-7.38 (m, 4H); HPLC Purity: 90.31%; Mass (M+1): 471.1.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(3,5-dichlorophenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 11.03 (s, 1H), 9.03 (s, 1H), 8.49 (s, 1H),8.39 (s, 1H), 7.24-7.21 (m, 4H); HPLC Purity: 97.52%; Mass (M+1): 472.8.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(3-ethylphenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.38 (s, 1H), 9.02 (s, 1H), 8.84 (s, 1H),8.37 (d, 2H), 7.17 (m, 1H), 7.01 (m, 2H), 6.86 (d, 1H), 2.52 (q, 2H),1.13 (t, 3H); HPLC Purity: 93.13%; Mass (M+1): 431.1.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(3-cyanophenyl)-1H-imidazole-5-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.99 (s, 1H), 9.04 (s, 1H), 8.87 (s, 1H),8.44 (s, 1H), 8.38 (s, 1H), 7.51 (m, 4H); HPLC Purity: 90.19%; Mass(M+1): 428.1.

General Procedure for Compound 3&3′: In a two neck RB flask, compound-2(3.0 g, 0.02 mole), pyrrole (2.69 g, 0.04 mole) and CS₂CO₃ (19.68 g,0.06 mole) were charged in toluene (60 mL) under N₂ atmosphere. BINAP(620 mg, 0.00099 mole) and Pd(OAc)₂ (990 mg, 0.004 mole) were then addedto the reaction mixture under N₂ atmosphere and stirred at 80° C. for3h. The progress of the reaction was monitored by TLC. After completionof SM, the reaction mixture was filtered through celite and quenchedwith water (5 mL). The organic layer was separated, dried over Na₂SO₄and concentrated under reduced pressure. The crude product was subjectedto silica gel column chromatography (silica gel 60-120, 1:9, ethylacetate/hexane) to separate the sulfonyl chlorides 3 & 3′ as viscous oil(compound-3: 1.1 g 30.55% and compound-3 ′: 684 mg 19.0%). Overall49.55% yield obtained.

¹H NMR (500 MHz, DMSO-d₆) δ: 8.53 (d, 1H), 7.54 (d, 2H), 7.15 (d, 1H),6.41 (d, 2H); Mass (M+1): 180.

General Procedure for Compound 4: In a single neck RB flask, tocompound-3 (100 mg, 0.59 mmol) was added chlorosulfonic acid (690 mg,5.9 mmol) slowly over a period of 15 min at 0° C. The resulting mixturewas stirred for 3 h at room temperature. After completion of SM, thereaction mixture was quenched with water (3 mL) and extracted with DCM(2×5 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated under reduced pressure. The crude compound was washed withpentane and dried under vacuum to provide the desired compound-4 as asolid (60 mg, 38.7%).

General Procedure for Compound 6: In a two neck RB flask, compound-5(9.5 mg, 0.16 mmol) and pyridine (0.15 mL) were taken and stirred for 10minutes at 0° C. Compound-4 (30 mg, 0.1 mmol) was then added at 0° C.and the resulting mixture was stirred for 3 h at room temperature. Aftercompletion of the reaction, HCl solution (6N, 4 mL) was added andextracted with DCM (2×5 mL). The combined organic layers were washedwith brine (1×20 mL), dried over anhydrous Na₂SO₄ and concentrated underreduced pressure. The crude product was purified by columnchromatography (silica gel 60-120, 2:8, ethyl acetate/hexane) to providethe desired compound-6 as a solid (20.4 mg, 68.1%).

¹H NMR (500 MHz, DMSO-d₆) δ: 8.81 (s, 1H), 8.19 (s, 1H), 7.97 (s, 1H),7.81 (s, 1H), 6.81 (s, 2H), 6.69 (s, 1H), 6.58 (s, 1H), 2.19 (s, 6H);HPLC Purity: 92.07%; Mass (M+1): 362.9.

General Procedure for Compound 7: In a single neck RB flask, tocompound-3′ (100 mg, 0.59 mmol) was added chlorosulfonic acid (690 mg,5.9 mmol) over a period of 15 min at 0° C. The resulting mixture wasthen stirred for 3 h at room temperature. Upon completion of SM (asindicated by TLC), the reaction mixture was quenched with water (3 mL)and extracted with DCM (2×5 mL). The combined organic layers were driedover Na₂SO₄ and concentrated under reduced pressure. The crude compoundwas washed with pentane and dried under vacuum to provide the desiredcompound-7 as a solid (35 mg, 22% yield).

General Procedure for Compound 8: In a two neck RB flask, compound-5(14.9 mg, 0.12 mmol) and pyridine (0.1 mL) were transferred and stirredfor 10 minutes at 0° C. Compound-7 (20 mg, 0.08 mmol) was then added at0° C. The resulting mixture was stirred for 3 h at room temperature.After completion of the reaction, HCl (6N, 4 mL) was added and extractedwith DCM (2×5 mL). The combined organic layers were washed with brine(1×20 mL), dried over Na₂SO₄ and concentrated under reduced pressure.The crude product was purified by column chromatography (silica gel60-120, 2:8, ethyl acetate/hexane) to provide the desired compound-8 asa solid (18.62 mg, 71% yield).

¹H NMR (500 MHz, DMSO-d₆) δ: 10.0 (s, 1H), 8.81 (s, 1H), 8.01 (s, 1H),7.78 (s, 1H), 7.63 (s, 1H), 6.82 (s, 2H), 6.68 (s, 1H), 6.54 (s, 1H),2.19 (s, 6H); HPLC Purity: 98.01%; Mass (M+1): 363.

1-(4-chloropyrimidin-2-yl)-N-(3,5-dimethylphenyl)-1H-pyrrole-2-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.0 (s, 1H), 8.81 (s, 1H), 8.01 (s, 1H),7.78 (s, 1H), 7.63 (s, 1H), 6.82 (s, 2H), 6.68 (s, 1H), 6.54 (s, 1H),2.19 (s, 6H); HPLC Purity: 98.01%; Mass (M+1): 363.

1-(2-chloropyrimidin-4-yl)-N-(3,5-dimethylphenyl)-1H-pyrrole-2-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 8.81 (s, 1H), 8.19 (s, 1H), 7.97 (s, 1H),7.81 (s, 1H), 6.81 (s, 2H), 6.69 (s, 1H), 6.58 (s, 1H), 2.19 (s, 6H);HPLC Purity: 92.07%; Mass (M+1): 362.9.

N-(4-chlorophenyl)-1-(2-chloropyrimidin-2-yl)-1H-pyrrole-2-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.34 (s, 1H), 8.82 (s, 1H), 8.23 (s, 1H),8.07 (s, 1H), 7.37 (d, 2H), 7.08 (d, 2H), 6.57 (s, 1H); HPLC Purity:96.74%; Mass (M+1): 369.

N-(4-chlorophenyl)-1-(4-chloropyrimidin-2-yl)-1H-pyrrole-2-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.29 (s, 1H), 8.90 (s, 1H), 8.02 (s, 1H),7.77 (s, 1H), 7.64 (s, 1H), 7.33 (d, 2H), 7.18 (d, 2H), 6.54 (s, 1H);HPLC Purity: 91.17%; Mass (M+1): 368.9.

N-(4-chlorophenyl)-1-(pyrimidin-2-yl)-1H-pyrrole-2-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.27 (s, 1H), 8.83 (d, 2H), 8.07 (s, 1H),7.79 (s, 1H), 7.48 (d, 1H), 7.32 (d, 2H), 7.18 (d, 2H), 6.53 (s, 1H);HPLC Purity: 97.05%; Mass (M+1): 334.9.

N-(3,5-dimethylphenyl)-1-(pyrimidin-2-yl)-1H-pyrrole-2-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 9.88 (s, 1H), 8.84 (d, 2H), 8.04 (s, 1H),7.78 (s, 1H), 7.43 (d, 1H), 6.79 (s, 2H), 6.63 (s, 1H), 6.49 (s, 1H);HPLC Purity: 97.48%; Mass (M+1): 329.

General Procedure for Compound 2: In a single neck RB flaskchlorosulfonic acid (9.7 ml, 145 mmol) was added slowly over a period of15 minutes to pyrazole (2.0 g, 29.0 mmol) at 0° C. The resulting mixturewas then heated at 100° C. for 3 h. Upon completion, the reactionmixture was quenched with ice cold water (25 mL) and extracted with DCM(3×40 mL). The combined organic layers were dried over Na₂SO₄ andconcentrated under reduced pressure. The desired product 2 was obtainedas a solid (4.8 g, 63% yield).

¹H NMR (500 MHz, CDCl3) δ: 8.19 (s, 2H).

General Procedure for Compound 3: To a stirred solution of compound-2(60 mg, 0.496 mmol) in DCM (10 mL) under a nitrogen atmosphere, pyridinewas added (0.1 mL, 0.99 mmol) and stirred at room temperature for 15minutes. The reaction mixture was then cooled to 0° C. followed by thedropwise addition of compound-2 solution (91.1 mg, 0.545 mmol in 4 mL ofDCM). The resulting mixture was stirred for 2 h at room temperature.After completion of the reaction, 0.5N HCl solution was added andextracted with DCM (2×10 mL). The combined organic layers were washedwith brine (1×20 mL), dried over Na₂SO₄ and concentrated under reducedpressure to furnish the desired compound-4 as a solid (120 mg, 97%yield).

¹H NMR (500 MHz, CDCl₃) δ: 10.4 (bs, 1H), 7.81 (s, 2H), 6.79 (s, 1H),6.72 (s, 2H), 6.47 (s, 1H), 2.23 (s, 6H); Mass (M+1): 252.

General Procedure for Compound 4: In a two neck RB flask, compound-3 (50mg, 0.2 mmol), compound-C (0.03 ml, 0.29 mmol) and K₂CO₃ (83 mg, 0.6mmol) were charged in DMF (5 mL) under N₂ atmosphere and stirred at 90°C. for 3 h. The progress of the reaction was monitored by TLC. Aftercompletion of SM, the reaction mixture was quenched with water (15 mL)and extracted with ethyl acetate (2×20 mL). The combined organic layerswere dried over Na₂SO₄ and concentrated under reduced pressure. Thecrude product was purified by column chromatography (Silica gel 60-120,3:7, ethyl acetate/hexane) to give the desired product 4 as a solid (70mg, 81.3% yield).

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(3,5-dimethylphenyl)-1H-pyrazole-4-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.10 (s, 1H), 9.01 (s, 1H), 8.82 (s, 1H),8.75 (s, 1H), 8.15 (s, 1H), 6.81 (s, 2H), 6.77 (s, 1H), 2.19 (s, 6H);HPLC Purity: 94.73%; Mass (M+1): 431.1.

N-(4-methoxyphenyl)-1-(5-(trifluromethyl)pyridin-2-yl)-1H-pyrazole-4-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 9.98 (s, 1H), 8.97 (s, 1H), 8.81 (s, 1H),8.45 (d, 1H), 8.10 (d, 1H), 8.05 (s, 1H), 7.07 (d, 2H), 6.85 (d, 2H),3.68 (s, 3H); HPLC Purity: 99.35%.

N-(4-chlorophenyl)-1-(5-(trifluromethyl)pyridin-2-yl)-1H-pyrazole-4-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.51 (s, 1H), 8.97 (s, 2H), 8.46 (d, 1H),8.18 (s, 1H), 8.12 (d, 1H), 7.38 (d, 2H), 7.21 (d, 2H); HPLC Purity:99.60%; Mass (M+1): 403.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(4-chlorophenyl)-1H-pyrazole-4-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 10.49 (s, 1H), 9.01 (s, 1H), 8.83 (s, 1H),8.78 (s, 1H), 8.13 (s, 1H), 7.38 (d, 2H), 7.19 (d, 2H); HPLC Purity:99.52%; Mass (M+1): 437.

1-(3-chloro-5-(trifluromethyl)pyridin-2-yl)-N-(4-methoxyphenyl)-1H-pyrazole-4-sulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 9.98 (s, 1H), 9.01 (s, 1H), 8.81 (s, 1H),8.64 (s, 1H), 8.02 (s, 1H), 7.07 (d, 2H), 6.84 (d, 2H), 3.68 (s, 3H);HPLC Purity: 99.35%; Mass (M+1): 432.9.

General Procedure for Preparation of Compound A: A solution of mixtureof triethyl orthoformate (11.2 g, 75 mmol) and malononitrile (5.0 g, 75mmol) in acetic anhydride (15 mL) was heated to 90° C. for 12 h. Aftercompletion of reaction (TLC shows absent of S.M), the solvent (excessacetic anhydride and acetic acid) was removed under high vacuum. Theresidue (6.0 g, 65.2% of yield) was used in the next reaction withoutfurther purification.

¹H NMR (500 MHz, DMSO-d6) δ: 8.59 (s, 1H), 4.55 (m, 2H), 1.38 (m, 3H).

General Procedure for Compound 2: To a stirred solution of2-bromo-5-(trifluoromethyl)pyridine (compound-1) (1.0 g, 4.0 mmol) inethanol (5 mL), hydrazine hydrate (980 mg, 20 mmol) was added at roomtemperature and the reaction mixture was heated to 90° C. for 12 h.After completion of reaction (TLC shows absent of S.M), the solvent wasremoved under high vacuum. The residue was treated with water (30 mL)and extracted with ethyl acetate (2×50 mL). The organic layer was washedwith brine (50 mL), dried over sodium sulfate, filtered and concentratedto provide the desired compound-2 as a brown color solid. (700 mg, 90%yield).

¹H NMR (500 MHz, DMSO-d₆) δ: 8.42 (d, 2H), 8.25 (s, 1H), 5.0 (bs, 3H);Mass (M+1): 178.

General Procedure for Compound 3: A solution of2-hydrazinyl-5-(trifluoromethyl)pyridine (compound-2) (250 mg, 1.4 mmol)and 2-(ethoxymethylene)malononitrile (compound-A) (170 mg, 1.4 mmol) inethanol (10 mL) was heated to 90° C. for 12 h. After completion ofreaction (TLC shows absent of S.M), the solvent was removed under highvacuum. The residue was treated with water (30 mL) and extracted withethyl acetate (2×50 mL). The organic layer was washed with brine (50mL), dried over sodium sulphate, filtered and concentrated to providethe desired compound-3 as an off white color solid. (280 mg, 80% yield).

¹H NMR (500 MHz, DMSO-d₆) δ: 8.91 (s, 1H), 8.45 (d, 1H), 8.32 (s, 2H),8.15 (d, 1H), 8.0 (s, 1H); Mass (M+1): 254.

General Procedure for Compound 4: In a two neck RB flask, compound-3 (50mg, 0.1 mmoL) was taken in THF (4 mL) and cooled to 0° C. under nitrogenatmosphere. NaH (60% NaH, 5 mg, 0.1 mmol) was added to the reactionmixture and stirred for 10 min followed by the addition of solution of3,5-dimethylbenzene-1-sulphonyl chloride (44 mg, 0.2 mmoL) in THF (2 mL)at 0° C. The reaction mixture was then stirred for 30 min. at roomtemperature. After completion of the reaction, the reaction mixture wasquenched with ice cold water and extracted with ethyl acetate, and theorganic layer was washed with brine (10 mL), dried over sodium sulphate,filtered and concentrated to provide the desired compound-4 as a solid(60 mg, 81.2% yield).

¹H NMR (500 MHz, DMSO-d₆) δ: 8.83 (s, 1H), 8.28 (d, 1H), 8.17 (d, 1H),7.77 (s, 1H), 7.31 (s, 2H), 7.01 (s, 1H), 2.21 (s, 6H); HPLC Purity:98.81%; Mass (M+1): 422.

N-(4-cyano-1-(5-(trifluromethyl)pyridin-2-yl)-1H-pyrazole-3,5dimethylbenzenesulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 8.83 (s, 1H), 8.28 (d, 1H), 8.17 (d, 1H),7.77 (s, 1H), 7.31 (s, 2H), 7.01 (s, 1H), 2.21 (s, 6H); HPLC Purity:98.81%; Mass (M+1): 422.

4-chloro-N-(4-cyano-1-(5-(trifluromethyl)pyridin-2-yl)-1H-pyrazol-5-yl)benzenesulfonamide

¹H NMR (500 MHz, DMSO-d₆) δ: 8.88 (s, 1H), 8.31 (d, 1H), 8.14 (bs, 1H),7.73 (m, 3H), 7.41 (m, 2H); HPLC Purity: 97.80%; Mass (M+1): 428.

Having thus described several aspects of at least one embodiment of thisinvention, it is to be appreciated various alterations, modifications,and improvements will readily occur to those skilled in the art. Suchalterations, modifications, and improvements are intended to be part ofthis disclosure, and are intended to be within the spirit and scope ofthe invention. Accordingly, the foregoing description and drawings areby way of example only.

Having thus described several aspects of at least one embodiment of thisinvention, it is to be appreciated various alterations, modifications,and improvements will readily occur to those skilled in the art. Suchalterations, modifications, and improvements are intended to be part ofthis disclosure, and are intended to be within the spirit and scope ofthe invention. Accordingly, the foregoing description and drawings areby way of example only.

The invention claimed is:
 1. A compound of formula (V) or apharmaceutically acceptable salt thereof, wherein:

B, D and E are each independently selected from H and R³; each R¹ isindependently selected from hydrogen and halo; R⁴ is hydrogen, C₁₋₈alkyl, and aryl, substituted with n occurrences of R²; each R² isindependently halo or alkyl; each R³ is independently selected fromhalo, haloalkyl and —OR^(a); R⁵ is hydrogen or C₁₋₈ alkyl; R^(a) isindependently selected from alkyl, haloalkyl, optionally substitutedheteroaryl and optionally substituted heterocyclyl; n is
 1. 2. Acompound of formula (V) or a pharmaceutically acceptable salt thereof,wherein:

B, D and E are each independently selected from H and R³; each R¹ isindependently selected from hydrogen and halo; R⁴ is hydrogen, C₁₋₈alkyl, and aryl, substituted with n occurrences of R²; each R² isindependently halo or alkyl; or two R², together with the carbon atomsto which they are attached, form a 5-membered heterocyclic ring; each R³is independently selected from halo, haloalkyl and —OR^(a); R⁵ ishydrogen or C₁₋₈ alkyl; R^(a) is independently selected from alkyl,haloalkyl, optionally substituted heteroaryl and optionally substitutedheterocyclyl; and n is
 2. 3. The compound of claim 2 or apharmaceutically acceptable salt thereof, wherein the compound isselected from: